Contents PCR...3 PCR TaqMan...6 TaqMan / PCR / PCR...8 PCR RNA...10 Cells-to-CT Kit...12 Cell-to-CT Kit RT-PCR TaqMa

Transcription

1 real-time PCR PCR

2 Contents PCR...3 PCR TaqMan...6 TaqMan / PCR / PCR...8 PCR RNA...10 Cells-to-CT Kit...12 Cell-to-CT Kit RT-PCR TaqMan Fast...18 TaqMan...19 SYBR Green...23 TaqMan...24 TaqMan TaqMan TaqMan...29 RNAi...32 sirna...33 RNA sirna mirna...36 microrna...37 microrna...38 microrna...39 microrna...40 Non-coding RNA...42 microrna...43 SNP...44 SNP...45 SNP High Resolution Melt HRM TaqMan INDEX...63 INDEX...69

3 PCR PCR TaqMan SYBR Green RNA PureLink RNA Kit TRIzol Reagent TRIzol Plus RNA Purification System TaqMan Cells-to-CT Kit Ambion Single Cell-to-CT Kit MagMax RNAqueous Kit SYBR Green Cells-to-CT Kit Super Script VILO 16 High Capacity RNA-to-cDNA Kit 17 1 RT-PCR TaqMan RNA-to-CT 1-Step Kit TaqMan Fast Virus 1-Step Master Mix Power SYBR Green RNA-to-CT 1-Step Kit 15 PCR TaqMan Fast Advance Master Mix TaqMan Fast Universal PCR Master Mix TaqMan Universal Master Mix II TaqMan Gene Expression Master Mix TaqMan Multiplex Master Mix Power SYBR Green PCR Master Mix Fast SYBR Green Master Mix SYBR Select Master Mix TaqMan TaqMan Gene Expression Assays TaqMan Array Plate TaqMan Array Gene Signature 96-well Plate TaqMan Gene Signature Arrays TaqMan Custom Arrays RNAi sirna Silencer Select sirna TaqMan sirna Assays Ambion In Vivo sirna Lipofectamie RNAiMAX Invivofectamine 3.0 Reagent microrna mirvana mirna Isolation Kit mirvana PARIS Kit TaqMan mirna ABC Purification Kit microrna TaqMan MicroRNA RT Kit 39 Megaplex Primer Pools 39 PCR TaqMan Universal Master Mix II 19 TaqMan TaqMan MicroRNA Assays TaqMan Array MicroRNA Card TaqMan Pri-miRNA Assays TaqMan Non-coding RNA Assays microrna microrna mimic & inhibitor mirvana mirna mimic & inhibitors 43 Lipofectamine RNAiMAX Invivofectamine 3.0 Reagent SNP TaqMan Genotyping Master Mix 46 PCR TaqMan GTXpress Master Mix 46 TaqMan TaqMan SNP Genotyping Assays 45 1 SNP TaqMan Sample-to-SNP Kit 47 TaqMan TaqMan Mutation Detection Assays 48 TaqMan TaqMan Copy Number Assays 49 HRM PCR MeltDoctor HRM 50 TaqMan TaqMan Protein Expression Core Reagents Kit with Master Mix TaqMan Protein Assay Kit TaqMan Protein Assay Open Kit Protein Thermal Shift Dye Kit 55 Protein Thermal Shift Starter Kit 55 3

4 PCR Applied Biosystems TaqMan Applied Biosystems SYBR Green PCR TaqMan 5'- TaqMan TaqMan PCR TaqMan PCR 5 - TaqMan PCR 5'- TaqMan TaqMan PCR DNA 5'-3' TaqMan 1.Polymerization TaqMan R Q 5' 3' TaqMan Gene Expression Assays Strand displacement TaqMan 3.Cleavage DNA 5'- 3' TaqMan 5' 4.Polymerization completed PCR 1. TaqMan 4

5 SYBR Green SYBR Green SYBR Green 2 DNA SYBR Green I PCR 2 DNA PCR SYBR Green I PCR Tm PCR 2 DNA SYBR Green I Dye DNA SYBR Green AmpliTaq Gold DNA Polymerase 1.Reaction setup SYBR Green I Dye 2 DNA 2.Denaturation DNA SYBR Green I Dye DNA 3.Polymerization PCR 4.Polymerization completed SYBR Green I Dye 2 DNA PCR 2. SYBR Green 1. TaqMan SYBR Green TaqMan SYBR Green TaqMan 2 DNA SYBR Green I 3 PCR 2 TaqMan 2 PCR PCR TaqMan Gene Expression Assays PCR 5

6 TaqMan TaqMan TaqMan microrna mirna Small RNA SNP TaqMan TaqMan SNP c TaqMan PCR TaqMan Gene Expression Assay / FAM VIC 4 c 2,700 TaqMan Drug Metabolism Genotyping c Assay PharmaADME DME mirna non-coding RNA mirna non-codingrna ncrna mirna pri-mirna small RNA mirna ncrna TaqMan Protein Assay c c SNP TaqMan SNP 450 TaqMan SNP Genotyping Assay Custom TaqMan SNP Genotyping Assay TaqMan Copy Number Assay Custom Plus TaqMan Copy Number Assays Custom TaqMan Copy Number Assays 2. TaqMan TaqMan Gene 1 TaqMan MGB 24 Expression Assays cdna Custom TaqMan Gene TaqMan Gene Expression Assays 24 Expression Assays TaqMan Array Plates TaqMan Gene Expression Assays 96 5 Fast 25 TaqMan Array Gene Signature 96-Well Plate TaqMan Gene Signature Arrays TaqMan Custom Arrays TaqMan Array Human Endogenous Control Plate Custom Plus TaqMan RNA Assays 96 TaqMan Gene Expression Assays 384 TaqMan TaqMan 32 TaqMan Gene Expression Assays

7 mirna non-coding RNA Megaplex RT Primer Pools for TaqMan MicroRNA Assays TaqMan MicroRNA Assays TaqMan Array MicroRNA Card mirna cdna Multiplex RT 1 mirna TaqMan mirna mirna TaqMan MicroRNA Assays 384 TaqMan 8 TaqMan Non-coding RNA Assays TaqMan Non-coding RNA Assays non-coding 42 TaqMan Pri-miRNA Assays mirna primary mirna pri-mirna 42 SNP TaqMan Pre-Designed SNP Genotyping Assays 450 SNP SNP 2 TaqMan PCR 45 TaqMan Drug Metabolism Genotyping Assays TaqMan Copy Number Assays 221 2,700 Genotyping TaqMan CNV TaqMan CNVs TaqMan Copy Number Assays TaqMan Copy Number Reference Assays PCR Custom TaqMan Copy Number Assays & Custom Plus TaqMan Copy Number Assays TaqMan Copy Number Reference Assays TaqMan Mutation Detection Assays TaqMan Copy Number Assays CNVs TaqMan Copy Number Reference Assays TaqMan Copy Number Assays Custom TaqMan Copy Number Assays TaqMan Competitive Allele-Specific TaqMan PCR castpcr MGB TaqMan Protein Assay Open Kit / TaqMan Protein Assays Core Reagents Kit with Master Mix TaqMan Protein Assay TaqMan 5 TaqMan Custom Plating Service TaqMan Gene Expression Assays TaqMan MicroRNA Assays / / 96 Standard 96 Fast 384 TaqMan Gene Expression Assays Inventoried, Made-to-Order, and Custom 24 TaqMan MicroRNA Assays Inventoried, Made-to-Order, and Custom 40 TaqMan Non-coding RNA Assays Inventoried, Made-to-Order 42 TaqMan SNP Genotyping Assays Predesigned Custom and DME 45 TaqMan Copy Number Assays Predesigned Custom Plus or Custom 49 7

8 TaqMan TaqMan シングルチューブ 96 もしくは 384 ウェルプレート 384 ウェル TaqMan Array カード OpenArray プレート 1 アッセイより購入可能 フレキシブル どんなタイプのリアルタイム PCR 装置にも対応 中小規模のプロジェクトに最適 反応セットアップの簡便さと柔軟性の適度なバランス 96 および 384 ウェルタイプのすべての装置に対応 1 反応当たりのコストが低い 中規模から大規模なプロジェクトに最適 Quant Studio 12K Flex Quant Studio 7 もしくは 7900HT リアルタイム PCR システムの TaqMan Array システムに対応 1 反応当たりのコストを最小に抑え大規模プロジェクトに最適 究極のスループット Applied Biosystems Quant Studio 12K Flex OpenArray システムに対応 凍結乾燥した混合済みアッセイおよびマスターミックスのフォーマットが利用可能 PCR / PCR PCR StepOne PCR 46 3 Fast StepOnePlus QuantStudio Fast 30 3 VeriFlex Block QuantStudio 7 New PCR (96 96 Fast 384 TaqManR Array ) QuantStudio 12K Flex Fast 384 TaqMan Array OpenArray ,000 OptiFlex LED 21 microrna PCR QuantStudio 3D PCR 20,000 8

9 PCR PCR PCR PCR PCR QuantStudio 3D PCR PCR 1% Custom TaqMan SNP Genotyping Assays EGFR, KRAS, BRAF, PIK3CA, JAK2 TaqMan QuantStudio 3D PCR PCR PCR PCR DNA DNA 3. Custom TaqMan SNP Genotyping Assays KRAS G12V QuantStudio 3D PCR DNA 10%(A) 1%(B) 0.1%(C) D PCR %(A) 1.393%(B) 0.142%(C) PCR 1 1 DNA PCR PCR PCR PCR 1 / PCR XX / µl QuantStudio 3D PCR PCR QuantStudio 3D PCR ProFlex PCR 1 QS3D-PF 6,400,000 QuantStudio PCR ProFlex PCR QuantStudio 3D ChipLoader 2 QuantStudio 3D PCR Chips v2 12 A ,880 QuantStudio 3D PCR Master Mix v2, 1.5 ml 1.5 ml A ,000 QuantStudio 3D PCR Master Mix v2, 5.0 ml 5.0 ml A ,000 Custom TaqMan SNP Genotyping Assays Custom TaqMan SNP Genotyping Assays for Rare Mutation Analysis ,500 20x, Small scale Custom TaqMan SNP Genotyping Assays for Rare Mutation Analysis 40x, Medium scale ,900 TaqMan Gene Expression Assays 9

10 RNA RNA PureLink RNA Kit Invitrogen PureLink RNA Mini Kit RNA RNA 1,000µg 100µg DNase Set DNA RNA(µg) 1, PureLink RNA Mini Kit Competitor s midi kit Competitor s mini kit Tissue(mg) 4. RNA PureLink RNA Mini Kit RNA PureLink RNA Mini Kit PureLink RNA Mini Kit , A 32, ,600 PureLink RNA Micro Kit ,900 PureLink Homogenizer ,600 PureLink DNase Set ,000 RNA TRIzol Reagent Invitrogen TRIzol Reagent DNA RNA TRIzol LS Reagent RNA TRIzol Reagent 100 L ,600 RNA TRIzol Plus RNA Purification System Invitrogen TRIzol Plus RNA Purification System Purelink RNA Mini Kit No.1 TRIzol 45 RNA TRIzol RNA TRIzol Plus RNA Purification System TRIzol Reagent 100 ml PureLink RNA Mini Kit A 200 L , L SP 178,000 TRIzol LS Reagent 100 L , L , , RNA TRIzol Reagent PureLink Kits TRIzol Plus RNA Purification System MagMAX Kits TRIzol mg ml 200 mg mg mg

11 RNA MagMax RNA RNA Applied Biosystems MagMax RNA MagMAX Total Nucleic Acid Isolation Kit RNA 100 AM ,800 MagMAX-96 Blood RNA Isolation Kit 96 AM ,800 MagMAX-96 Total RNA Isolation Kit 96 AM ,700 MagMAX Viral RNA Isolation Kit 50 AM ,700 MagMAX-96 Viral RNA Isolation Kit 96 AM ,800 RNA Single Place Magnetic Stand 1 AM ,900 Magnetic Stand-96 1 AM , well Magnetic-Ring Stand 1 AM ,000 RNA RNAlater Solution Applied Biosystems RNAlater Solution RNA RNAlater Solution RNAlater Solution 100 ml AM , ml AM , ml AM , ml AM , ml AM ,400 RNA RNAlater -ICE Applied Biosystems RNAlater -ICE RNA RNAlater -ICE -20 RNA RNAlater -ICE 25 ml AM ,800 RNAqueous Kit Applied Biosystems RNAqueous Kit RNA 20 RNA DNA RNA RNAqueous-4PCR Kit RNAqueous Kit 50 AM ,900 RNAqueous Midi Kit 15 AM ,700 RNAqueous-4PCR Kit 30 AM ,000 RNAqueous-96 Kit 192 AM ,400 RNAqueous-96 Automated Kit 4 96 AM ,600 RNAqueous-Micro Kit 50 AM ,600 DNA TURBO DNA-free Kit Applied Biosystems TURBO DNA-free Kit DNA TURBO DNase TURBO DNase DNase I DNA DNase I 6 RT-PCR false positive DNA TURBO DNA-free Kit 50 AM ,300 RNA THE RNA Storage Solution 10 1 ml AM7000 8, ml AM7001 8,400 TE Buffer ph ml AM9860 8, ml AM9861 8, mm EDTA 50 ml AM9912 8,400 11

12 Cells-to-CT Kit RNA Cells-to-CT Kit Applied Biosystems Cells-to-CT Kit RNA PCR RNA Cells-to-CT Kit sirna microrna mirna TaqMan Gene Expression Cells-to-CT Kit TaqMan Fast Cells-to-CT Kit Cells-to-CT 1-Step TaqMan Kit Power SYBR Green Cells-to-CT Kit Cells-to-CT 1-Step Power SYBR Green Kit Fast SYBR Green Cells-to-CT Kit gdna PCR DNase ,000 RNA TaqMan PreAmp Cells-to-CT Kit mirna TaqMan MicroRNA Cells-to-CT Kit 細胞溶解 (7 分 ) 逆転写反応リアルタイム解析 2 Step 5. 1 Step RNA 6. Cells-to-CT Kit PCR Cells-to-CT CTs HeLa RNA Cells-to-CT Kit 128 TaqMan Gene Expression Assays Applied Biosystems 7900HT PCR Cells-to-CT Kit Ct RNA Ct Purified RNA C T s 12

13 TaqMan TaqMan Gene Expression Cells-to-CT Kit Applied Biosystems TaqMan Gene Expression Cells-to-CT Kit RNA DNA cdna PCR TaqMan Gene Expression Master Mix TaqMan Gene Expression Cells-to-CT Kit * , AM , AM ,100 TaqMan Cells-to-CT Control Kit/Stop Solution Applied Biosystems TaqMan Cells-to-CT Control Kit TaqMan Gene Expression Cells-to-CT Kit PCR TaqMan Cells-to-CT Control Kit ,500 Cells-to-CT Stop Solution 1 ml ,700 TaqMan Fast Cells-to-CT Kit Applied Biosystems TaqMan Fast Cells-to-CT Kit RNA Fast PCR PCR TaqMan Fast Universal PCR Master Mix TaqMan Fast Cells-to-CT Kit ,500 * Cells-to-CT 1-Step TaqMan Kit TaqMan 1-Step RT-PCR RT-PCR Applied Biosystems Cellsto-CT 1-Step TaqMan Kit Cells-to-CT Fast 1 RT-PCR Cells-to-CT 1-Step TaqMan Kit 20 A , A , A ,200 SYBR Green Power SYBR Green / Fast SYBR Green Cells-to-CT Kit Applied Biosystems SYBR Green Cells-to-CT Kits SYBR Green DNA PCR Power SYBR Green Master Mix Fast SYBR Green Master Mix Power SYBR Green Cells-to-CT Kit Power SYBR Green Cells-to-CT Kit , , ,500 * Fast SYBR Green Cells-to-CT Kit Fast SYBR Green Cells-to-CT Kit , ,000 * SYBR Green Cells-to-CT Control Kit SYBR Green Cells-to-CT Control Kit ,500 Cells-to-CT 1-Step Power SYBR Green Kit Applied Biosystems Cells-to-CT 1-Step Power SYBR Green Kit RT-PCR 1 Power SYBR Green 1-Step RT-PCR Cells-to-CT 1-Step 20 A ,300 Power SYBR Green Kit 100 A , A ,300 mirna TaqMan MicroRNA Cells-to-CT Kit Applied Biosystems TaqMan MicroRNA Cells-to-CT Kit mirna mirna TaqMan MicroRNA Cells-to-CT Kit ,400 * TaqMan PreAmp Cells-to-CT Kit Applied Biosystems TaqMan PreAmp Cells-to-CT Kit TaqMan Gene Expression Cells-to-CT Kit TaqMan Gene Expression Master Mix cdna TaqMan PreAmp Master Mix TaqMan PreAmp Cells-to-CT Kit ,100 * 13

14 Cell-to-CT Kit Ambion Single Cell-to-CT Kit Invitrogen Ambion Single Cell-to-CT Kit 1 10 RNA 5 Pre-Amplification PCR RNA Ambion Single Cell Lysis Kit RNA Invitrogen Ambion Single Cell Lysis Kit RNA RNA PCR Single Cell-to-CT Kit 28 CT µL の Single Cell Solution に 1 10 細胞を加えます 室温で 5 分インキュベートします µL の Stop Solution にを加えます 室温で 2 分インキュベートします Cell Equivalent Single Cells Cells Ambion Single Cell-to-CT Kit , ,090,400 Ambion Single Cell Lysis Kit ,400 * 7. Single Cell-to-CT Kit CT T DNA Cells-to-CpG Bisulfite Conversion Kit Applied Biosystems Cells-to-CpG Bisulfite Conversion Kit DNA FFPE DNA Single Cell Lysis Buffer PBS / 5 3. PBS RT cdna 14 Pre-Amplification B2M GAPDH HPRT1 TGFB1 5 Ct Cells-to-CpG Bisulfite Conversion ,000 Kit ,000 Cells-to-CpG Bisulfite Conversion ,500 & Quantitation Control Kit Cells-to-CpG Methylated and Unmethylated gdna Control Kit 50 µl ,000 14

15 1 RT-PCR TaqMan RNA-to-CT 1-Step Kit Applied Biosystems TaqMan RNA-to-CT 1-Step Kit 1 TaqMan 1 1 TaqMan RNA TaqMan Fast Virus 1-Step Master Mix 21 1 AT GC TaqMan Gene Expression Assays 24 TaqMan RNA-to-CT 1-Step Kit, ,200 1 ml TaqMan RNA-to-CT 1-Step Kit, ,400 5 ml TaqMan RNA-to-CT 1-Step Kit, 2, ,000 5 ml 10 TaqMan RNA-to-CT 1-Step Kit TaqMan RNA-to-CT 1-Step Kit Power SYBR Green RNA-to-CT 1-Step Kit Applied Biosystems Power SYBR Green RNA-to-CT 1-Step Kit PCR 1 SYBR Green Rn Cycle Power SYBR Green RNA-to-CT ,200 1-Step Kit, 1mL Power SYBR Green RNA-to-CT ,400 1-Step Kit, 5mL Power SYBR Green RNA-to-CT Step Kit Power SYBR Green RNA-to-CT 1-Step Kit Derivative Reporter (-Rn) Temperature (ºC) 10. Power SYBR Green RNA-to-CT 1-Step Kit GAPDH RNA BOSC23 Applied Biosystems StepOnePlus PCR 3 15

16 逆転写 PCR SuperScript VILO Invitrogen SuperScript VILO Variable Input Linear Output SuperScript RT PCR mrna cdna RNA SuperScript VILO Master Mix 1 VILO SuperScript VILO PCR cdna PCR 1pg ~ 2.5µg の広範囲で高い直線性 逆転写 高次構造の RNA テン レー cdna 合成 RNA が高い て逆転写 合 逆転写 性 テッ で の cpm , , ,965 0 S 製品 VILO A 製品 11. cdna 32 P-dCTP cdna S A SuperScript VILO VILO A 9 従来高次構造により逆転写が停止 VILO ヘルパータンパク質等により高次構造がほぐれて cdna 合成 13. cdna PCR TaqMan TfR Target SuperScript VILO cdna Synthesis Kit SuperScript VILO cdna Synthesis Kit , ,400 SuperScript VILO Master Mix 他の製品と比較して最大のダイナミックレンジ SuperScript VILO Master Mix , , ,100 RNA cdna 合成 RNA cdna 合成 12. RNA cdna VILO RNA cdna 4. High Capacity cdna High Capacity SuperScript VILO SuperScript VILO Reverse Transcriptase Kit RNA-to-cDNA Kit cdna Synthesis Kit Master Mix MultiScribe MultiScribe Superscript III Superscript III 37 C 37 C 42 C 42 C (Total RNA) 20 ng-2 µg 20 ng-2 µg 1 pg-2.5 µg 1 pg-2.5 µg RNase H

17 High Capacity RNA-to-cDNA Kit Applied Biosystems High Capacity RNA-to-cDNA Kit PCR cdna RNA PCR High Capacity RNA-to-cDNA Kit RNA cdna 2 High Capacity cdna Reverse Transcription Kit High Capacity cdna Reverse ,300 Transcription Kit 1, ,600 High Capacity cdna Reverse ,700 Transcription Kit, with RNase Inhibitor 1, ,800 High Capacity cdna Reverse Transcription Kit RNase Inhibitor, 20 units/µl 100 N ,400 *20 µl µg RNA 2 RT-PCR High Capacity RNA-to-cDNA Kit ,300 High Capacity RNA-to-cDNA Kit High Capacity RNA-to-cDNA Kit *20 µl TaqMan Reverse Transcriation Reagents TaqMan Reverse 200 N ,600 Transcription Reagents 10-Pack, TaqMan Reverse Transcription Reagents 2, ,900 / / MultiScribe Reverse Transcriptase 5,000 units ,700 RNase Inhibitor, 20 units/µl 2,000 units N ,400 Random Hexamers, 5 nmoles 0.1 ml N ,500 Oligo d(t) 16 Primers, 5 nmoles 0.1 ml N ,500 GeneAmp dntp Blend, 10 mm 1 ml N ,200 GeneAmp dntp Blend, 12.5 mm 100 N ,100 with dutp AmpErase Uracil N-glycosylase 100 units N ,900 UNG 20 % Glycerol Solution, 100 ml ,500 Molecular Biology Grade TaqMan Control Total RNA Human 100 µl , ng/µl TaqMan Control Genomic DNA Human 10 ng/µl, µl ,500 17

18 TaqMan Fast Fast 10 PCR Fast Fast 40 Applied Biosystems TaqMan Fast Advanced Master Mix Fast Fast PCR Fast Fast PCR ΔRn Cycle Number TaqMan Fast Advanced Master Mix 40 TaqMan Fast Universal PCR Master Mix Applied Biosystems TaqMan Fast Universal PCR Master Mix TaqMan Fast PCR Fast PCR Fast PCR Fast AmpliTaq Fast DNA Polymerase 72 PCR TaqMan Assays TaqMan Array Fast Fast Fast TaqMan Fast Universal PCR ,000 Master Mix 2X, No AmpErase UNG 2-Pack, TaqMan Fast Universal PCR ,700 Master Mix 2X, No AmpErase UNG 5-Pack, TaqMan Fast Universal PCR 1, ,100 Master Mix 2X, No AmpErase UNG 10-Pack, TaqMan Fast Universal PCR 2, ,400 Master Mix 2X, No AmpErase UNG TaqMan Fast Universal PCR 5, ,500 Master Mix 50 ml TaqMan Fast Universal PCR Master Mix *20 µl TaqMan Fast Advanced Master Mix, ,800 1 ml TaqMan Fast Advanced Master Mix, ,900 5 ml TaqMan Fast Advanced Master Mix, 1, , ml TaqMan Fast Advanced Master Mix, 2, , ml TaqMan Fast Advanced Master Mix, 5, , ml TaqMan Fast Advanced Master Mix, 5, , ml *20 µl 18

19 TaqMan 1 TaqMan Universal Master Mix II Applied Biosystems TaqMan Universal Master Mix II microrna mirna SNP CNV Applied Biosystems TaqMan Universal Master Mix II 1 RNase P TaqMan Universal Master Mix II PCR PCR 24 SNP mirna TaqMan Assays TaqMan Assays PCR UNG PCR TaqMan Universal Master Mix II, with UNG TaqMan Universal Master Mix II, ,800 with UNG 1 ml TaqMan Universal Master Mix II, ,900 with UNG 5 ml TaqMan Universal Master Mix II, ,100 with UNG 2 5 ml TaqMan Universal Master Mix II, 1, ,500 with UNG 5 5 ml TaqMan Universal Master Mix II, 2, ,400 with UNG 10 5 ml TaqMan Universal Master Mix II, with UNG 50 ml 2, ,000 TaqMan Universal Master Mix II, no UNG TaqMan Universal Master Mix II, ,800 no UNG 1 ml TaqMan Universal Master Mix II, ,200 no UNG 5 ml TaqMan Universal Master Mix II, ,900 no UNG 2 5 ml TaqMan Universal Master Mix II, 1, ,400 no UNG 5 5 ml TaqMan Universal Master Mix II, 2, ,000 no UNG 10 5 ml TaqMan Universal Master Mix II, no UNG 50 ml 2, ,800 *50 µl PCR 24 PCR TaqMan Universal Master Mix II PCR PCR

20 TaqMan TaqMan Gene Expression Master Mix Applied Biosystems TaqMan Gene Expression Master Mix PCR GC AT PCR TaqMan Universal PCR Master Mix 72 TaqMan Gene Expression ,800 Master Mix, 1 ml TaqMan Gene Expression ,900 Master Mix, 5 ml TaqMan Gene Expression ,100 Master Mix, 2 5 ml TaqMan Gene Expression 1, ,500 Master Mix, 5 5 ml TaqMan Gene Expression 2, ,400 Master Mix, 10 5 ml TaqMan Gene Expression 2, ,000 Master Mix, 50 ml TaqMan Gene Expression Master Mix TaqMan Gene Expression Master Mix *50 µl RNA PCR Water Nuclease free Water Nuclease-free Water 500 ml AM ,000 1,000 ml AM , ml AM , ml AM9938 6, ml AM ,900 TaqMan Universal PCR Master Mix Applied Biosystems TaqMan Universal PCR Master Mix TaqMan 5' PCR Applied Biosystems AmpliTaq Gold DNA Polymerase TaqMan Universal PCR Master Mix ,400 2-Pack, ,500 TaqMan Universal PCR Master Mix Pack, 1, ,800 TaqMan Universal PCR Master Mix Pack, 2, ,100 TaqMan Universal PCR Master Mix TaqMan Universal PCR Master Mix, 10-Pack 5 ml 10 2, ,100 TaqMan Universal PCR Master Mix, 50 2, ,900 ml TaqMan Universal PCR Master Mix *50 µl TaqMan Universal PCR Master Mix, No AmpErase UNG Applied Biosystems TaqMan Universal PCR Master Mix, No AmpErase UNG AmpErase UNG TaqMan 5' PCR AmpliTaq Gold DNA Polymerase TaqMan Universal PCR Master Mix, ,900 No AmpErase UNG TaqMan Universal PCR Master Mix, ,400 No AmpErase UNG, 2-Pack TaqMan Universal PCR Master Mix, 1, ,400 No AmpErase UNG, 5-Pack TaqMan Universal PCR Master Mix, 2, ,700 No AmpErase UNG, 10-Pack TaqMan Universal PCR Master Mix, 2, ,500 No AmpErase UNG, 50 ml TaqMan Universal PCR Master Mix *50 µl 20

21 PCR TaqMan Fast Virus 1-Step Master Mix PCR RNA DNA 1 4 RNA / DNA PCR Fast RT-PCR PCR TaqMan PreAmp Master Mix Applied Biosystems TaqMan PreAmp Master Mix cdna PrePCR PrePCR TaqMan Gene Expression Assays 100 1ng cdna 200 PCR TaqMan PreAmp Master Mix ,400 TaqMan PreAmp Master Mix Kit ,600 PreAmp Master Mix ( ) TaqMan Gene Expression Master Mix, 5 ml ( ) PCR Applied Biosystems TaqMan Fast Virus 1-Step Master Mix TaqMan Reagents Starter Kit TaqMan Reagents Starter Kit 1 Kit ,400 TaqMan Universal Master Mix, No UNG 1.25 ml TaqMan Gene Expression Assay, Eukaryotic 18S rrna, 20X 0.3 ml Human Raji cdna, 25 ng/µl 0.05 ml Fast PCR TaqMan Fast Virus 1-Step Master Mix PCR 4 Fast 一般的な 2 ステップ反応 TaqMan EZ RT-PCR Core Reagents TaqMan RNA-to-C T 1-Step Kit TaqMan Fast Virus 1-Step Master Mix リアルタイム PCR の所要時間 ( 分 ) Fast TaqMan Fast Virus 1-Step Master Mix 1 1 TaqMan Fast Virus 1-Step Master Mix TaqMan Fast Virus 1-Step Master Mix, 1 ml TaqMan Fast Virus 1-Step Master Mix, 5 1 ml TaqMan Fast Virus 1-Step Master Mix, 10 ml *20 µl ,800 1, ,800 2, ,500 21

22 TaqMan TaqMan Multiplex Master Mix Applied Biosystems TaqMan Multiplex Master Mix Fast PCR 4 TaqMan Multiplex Master Mix AmpliTaq Fast DNA Polymerase Uracil-N Glycosylase UNG dutp dntp MUSTANG PURPLE 2X 4 SNP Fast PCR Standard PCR Fast PCR UNG UDG TaqMan Multiplex Master Mix 1 ml ,100 5 ml ,100 2 x 5 ml ,400 5 x 5 ml , x 5 ml , ml ,700 TaqMan GAPDH Assay, 250 reactions ,000 JUN dye/qsy probe TaqMan GAPDH Assay, 250 reactions ,000 JUN dye/qsy probe, primer-limited TaqMan RNase P Assay, 250 reactions ,000 ABY dye/qsy probe TaqMan RNase P Assay, ABY dye/qsy probe, primer-limited 250 reactions ,000 TaqMan QSY TaqMan QSY 6,000 pmol ,500 20,000 pmol ,200 50,000 pmol ,300 QSY A Rn B Rn C Rn D Rn Amplification plot Cycle Amplification plot Cycle Amplification plot Cycle Amplification plot Cycle 17. A EGFR FAM B BRCA1 VIC C ESR1 JUN D RNaseP ABY QuantStudio 7 PCR TaqMan Multiplex Master Mix cdna 10 µl 20,000 pg 2 pg 4 4-plex Ct 4-plex PCR EGFR 96.09% 4-plex 96.39% BRCA % 4-plex 94.93% ESR % 4-plex 95.81% RNaseP 96.91% 4-plex 98.1% TaqMan QSY Applied Biosystems TaqMan QSY 4 3 MGB QSY 22

23 SYBR Green Power SYBR Green PCR Master Mix Applied Biosystems Power SYBR Green PCR Master Mix 2 DNA cdna RNA PCR SYBR Green PCR 2 Power SYBR Green PCR Master Mix, 1 ml Power SYBR Green PCR Master Mix, 5 ml 2-Pack, Power SYBR Green PCR Master Mix, 2 5 ml 5-Pack, Power SYBR Green PCR Master Mix, 5 5 ml 10-Pack, Power SYBR Green PCR Master Mix, 10 5 ml Power SYBR Green PCR Master Mix, 50 ml Power SYBR Green PCR Master Mix *50 µl , , ,400 1, ,300 2, ,000 2, , Fast SYBR Green Master Mix Applied Biosystems Fast SYBR Green Master Mix Fast PCR Fast PCR PCR Fast SYBR Green Master Mix, 1 ml ,500 Fast SYBR Green Master Mix, 5 ml ,800 2-Pack, Fast SYBR Green Master Mix, 2 5 ml 5-Pack, Fast SYBR Green Master Mix, 5 5 ml 10-Pack, Fast SYBR Green Master Mix, 10 5 ml 1, ,400 2, ,300 5, ,000 Fast SYBR Green Master Mix, 50 ml 5, ,000 Fast SYBR Green Master Mix Fast SYBR Green Master Mix *20 µl Delta Rn vs Cycle 1.0e+002 Delta Rn 1.0e e e e ng 5 ng 500 pg 50 pg 5 pg Cycle Number 19. Fast SYBR Master Mix SYBR Green PCR Master Mix Applied Biosystems SYBR Green PCR Master Mix SYBR Green PCR 2 CT Standard Curve pg Slope = pg 28 R 2 = pg ng ng Log Concentration SYBR Green PCR Master Mix, 1 ml ,500 SYBR Green PCR Master Mix, 5 ml ,500 SYBR Green PCR Master Mix, 50 ml 2, , Pack, SYBR Green PCR Master Mix, 10 5 ml SYBR Green PCR Master Mix *50 µl 2, , Human gdna -actin 23

24 TaqMan TaqMan Gene Expression Assays 5. TaqMan Gene Expression Assays Applied Biosystems TaqMan Gene Expression Assays PCR & 130 Exon RNAi FAM TaqMan Gene Expression Assays, ,500 Inventoried TaqMan Gene Expression Assays, ,700 Small, Made-to-Order TaqMan Gene Expression Assays, ,000 Medium, Made-to-Order TaqMan Gene Expression Assays, 2, ,900 Large, Made-to-Order VIC TaqMan Gene Expression Assays, ,700 Small_VIC, Made-to-Order TaqMan Gene Expression Assays, ,000 Medium_VIC, Made-to-Order TaqMan Gene Expression Assays, Large_VIC, Made-to-Order 2, ,900 Primer Limited TaqMan Assays VIC TaqMan Gene Expression Assays, ,700 Small_VIC, PL Made-to-Order TaqMan Gene Expression Assays, ,000 Medium_VIC, PL Made-to-Order TaqMan Gene Expression Assays, Large_VIC, PL Made-to-Order 2, ,900 *20 µl Inventoried Made-to-Order 60 Species Number of Assays H. sapiens (Human) 212,207 M. musculus (Mouse) 185,566 R. norvegicus (Rat) 151,955 O. sativa (Rice) 99,822 A. thaliana (Arabidopsis) 97,879 C. elegans (Nematode) 92,687 B. taurus (Cow) 67,104 M. mulatta (Rhesus Monkey) 66,480 D. rerio (Zebrafish) 63,712 X. tropicalis (Frog) 56,764 C. familiaris (Dog) 55,202 G. gallus (Chicken) 48,432 Species Number of Assays D. melanogaster (Fruit Fly) 41,607 Z. mays (Sweet Corn) 38,446 S. scrofa (Pig) 9,868 S. pombe (Fission Yeast) 6,538 O. cuniculus (Rabbit) 5,924 S. cerevisiae (Baker's Yeast) 5,525 G. max (Soybean) 3,456 E. caballus (Horse) 2,970 C. porcellus (Guinea Pig) 2,037 V. vinifera (Grape) 965 T. aestivum (Wheat) 760 Total Number of Assays 1,315,906 Custom TaqMan Gene Expression Assays Applied Biosystems Custom TaqMan Gene Expression Assays TaqMan Gene Expression Assays TaqMan Gene Expression Assays TaqMan Custom TaqMan Gene 50 µl ,700 Expression Assays, S-scale 20 µl 360 Custom TaqMan Gene Expression Assays, M-scale Custom TaqMan Gene Expression Assays, L-scale 50 µl µl µl 1, µl 2, , ,600 Custom Plus TaqMan RNA Assays Applied Biosystems Custom Plus TaqMan RNA Assays TaqMan Gene Experssion Assays Custom Plus TaqMan RNA Assays Custom Plus TaqMan RNA Assay, ,700 Small 20 Custom Plus TaqMan RNA Assay, Medium ,100 Custom Plus TaqMan RNA Assay, Large 60 2, ,600 24

25 TaqMan 96 TaqMan Array Plate 96 - Fast Applied Biosystems TaqMan Array Plate TaqMan Gene Expression Assays Fast 10 TaqMan Gene Expression Assays 6 96 PCR TaqMan Standard : 7900HT / ViiA 7 / 7500 / 7300 / QuantStudio 96 Fast : 7900HT / ViiA 7 /7500Fast / StepOnePlus /Quant- Studio Fast96 60 TaqMan Array Plate ** * TaqMan Array Plates, Format 8 (Standard) TaqMan Array Plates, Format 8 (Fast) TaqMan Array Plates, Format 16 (Standard) TaqMan Array Plates, Format 16 (Fast) TaqMan Array Plates, Format 16 + Endogenous Controls (Standard) TaqMan Array Plates, Format 16 + Endogenous Controls (Fast) TaqMan Array Plates, Format 32 (Standard) TaqMan Array Plates, Format 32 (Fast) TaqMan Array Plates, Format 32 + Endogenous Controls (Standard) TaqMan Array Plates, Format 32 + Endogenous Controls (Fast) TaqMan Array Plates, Format 48 (Standard) TaqMan Array Plates, Format 48 (Fast) TaqMan Array Plates, Format 48 + Endogenous Controls (Standard) TaqMan Array Plates, Format 48 + Endogenous Controls (Fast) TaqMan Array Plates, Format 96 (Standard) TaqMan Array Plates, Format 96 (Fast) TaqMan Array Plates, Format 96 + Endogenous Controls (Standard) TaqMan Array Plates, Format 96 + Endogenous Controls (Fast) ,100 29,600 25, ,100 29,600 25, ,700 34,100 29, ,700 34,100 29, ,700 34,100 29, ,700 34,100 29, ,200 44,300 38, ,200 44,300 38, ,200 44,300 38, ,200 44,300 38, ,800 51,200 44, ,800 51,200 44, ,800 51,200 44, ,800 51,200 44, ,000 56,800 51, ,000 56,800 51, ,000 56,800 51, ,000 56,800 51,200 * ** Candidate Endogenous Control GAPDH HPRT1 GUSB 3 25

26 TaqMan 96 TaqMan Array Gene Signature 96-Well Plate Applied Biosystems TaqMan Array Gene Signature 96-Well Plate TaqMan Gene Expression Assays Standard Fast Human Alzheimer's Disease Human Angiogenesis Human Apoptosis Human Immune Response Human Inflammation Human Phosphodiesterase Human Protein Kinase Pathways Human Stem Cell Pluripotency Mouse Alzheimer's Disease Mouse Immune Response Mouse Stem Cell Pluripotency Rat Inflammation Rat Phosphodiesterase Human Androgens Human Chemokines Human CYP450 & other Oxygenases Human Diabetes Human Estrogens Human Heat Shock Proteins Human Hedgehog Pathway Human Hypoxia Human LDL Pathways Human Ligand Gated Ion Channels Human MAPK Pathways Human Neurotransmitters Human NFkB Pathway Human Osteogenesis Human TGF Pathway Human Tumor Metastasis Human Voltage-Gated Ion Channels Human WNT Pathway Human Induced Intracellular Signaling Human Activation of camp-dependent PKA Human HOX Genes Human Antigen Processing and Presentation by MHCS Human Apoptosis via Death Receptors Human Apoptotic Pathways Triggered By HIV Mouse ABC Transporters Human Nuclear Receptors Human Cell Surface Markers Human BMP Pathway ,700 Standard Fast Human BRCA1 Pathway Human Breast Cancer Regulations by Stathmin Human c-myc and Apoptosis Human camp Pathway Human Cardiomyocyte Differentiation by BMP Receptors Human Cell Cycle Control of Chromosomal Replication Human Cellular Apoptosis Pathway Human Drug Transporters Human Antioxidant Mechanisms Human Complement Pathway Human Colorectal Cancer Metastasis Human CREB Pathway Human Cyclins & Cell Cycle Regulation Human Cytokine Network Human Developmental Phases of Colorectal Cancer Human DNA Damage Induced Sigma Signaling Human DNA Methylation & Transcriptional Repression Human DNA Repair Mechanisms Human EGF Pathway Human Signal Transduction Pathways Human enos Signaling Human Erythropoietin Pathway Human Extracellular Matrix & Adhesion Molecules Human Factors for Cardiogenesis Human FAS Signaling Human FGF Pathway Human GPCR Pathway Human Guidance & Growth Cone Motility Human Transcription Factors (Non-Hox) Human Huntington Disease Pathway Human IGF1R Signaling Human IL-1 Pathway Human IL-10 Pathway Human IL-2 Active 8 & Quiescent T-Cell Human IL-2 Pathway Human AHR Pathway Human IL-3 Pathway Human IL-4 Pathway

27 Standard Fast Human IL-6 Pathway Human IL-9 Pathway Human ILK Signaling Human inos Signaling Human Insulin Receptor Pathway Human Interferon Pathway Human Intracellular Calcium Signaling Human JAK-STAT Pathway Human Growth Factors Human Hematopoiesis Human Melatonin Signaling Human Prion Pathway Human Molecular Mechanisms of Cancer Human NGF Pathway Human nnos Signaling at Neuronal Synapses Human nnos Signaling in Skeletal Muscle Human Notch Signaling Human ABC Transporters Human P53 Mediated Apoptosis Human P53 Signaling Human CMV & MAPK Pathways Human Patho Rheumatoid Arthritis Human PDGF Pathway Human PI3K Signaling Human PI3K Signaling in B-Lymphocyte Human PKA Signaling Human Cholera Infection Human RAS Pathway Human Pancreatic Adenocarcinoma Human Phagocytosis of Microbes Human RHO Family GTPases Human RHOA Pathway Human Pathogenesis of ALS Human Signaling in Gap Junctions Human NFAT & Cardiac Hypertrophy Human SMAD Signaling Network Human Sperm Motility Human T-Cell Receptor and CD3 Complex Human Telomere Extension by Telomerase Human Thrombin Signaling Human TNF Superfamily Pathway Human TNFR1 Pathway ,700 Standard Fast Human TNFR2 Pathway Human Toll Comparative Pathway Human Toll-Like Receptors Pathway Human Transcription of mrna Human Transcription of rrna Human Transcription of trna Human Transcriptional Regulatory Network in Embryonic Stem Cell Human Ubiquitin-Proteasome Dependent Proteolysis Human VEGF Pathway Human Maturity-Onset Diabetes of Young MODY Mouse Lipid Regulated Genes Rat Endogenous Control Mouse Endogenous Control Human Endogenous Control

28 TaqMan 384 TaqMan Gene Signature Arrays Applied Biosystems TaqMan Gene Signature Arrays TaqMan Human Human Immune Array ,000 Human Endogenous Control Array ,100 Human Protein Kinase Array ,000 Human GPCR Array ,700 Human ABC Transporter Array ,000 Human Apoptosis Array ,000 Human Phosphodiesterase Array ,000 Human Inflammation Array ,000 Human Angiogenesis Array ,000 Human Alzheimer's Array ,000 Human Nuclear Receptor Array ,000 Human Stem Cell Pluripotency Array ,000 TaqMan Array Human MicroRNA A Cards v ,000 TaqMan Arrays Human MicroRNA B Cards v ,000 TaqMan Arrays Human MicroRNA A+B Cards v ,800 Mouse Mouse Immune Array ,000 Mouse Endogenus Control Array ,100 Mouse GPCR Array ,700 Mouse Alzheimer's Array ,000 Mouse Stem Cell Pluripotency Array ,000 TaqMan Array Rodent MicroRNA A Cards v ,000 Rat Rat Endogenous Control Array ,100 Rat Phosphodiesterase Array ,000 Rat Inflammation Array ,000 Rat GPCR Array ,700 TaqMan Array Rodent MicroRNA A Cards v ,000 TaqMan Custom Arrays Micro Fluidic Card Applied Biosystems TaqMan Custom Arrays 7900HT Fast ViiA 7 QuantStudio PCR TaqMan Custom Arrays 7900HT ViiA 7 QuantStudio TaqMan Array Micro Fluidic Card 2 µl 384 TaqMan Gene Signature Arrays TaqMan Array, Format ,200 63,600 51,200 45,400 TaqMan Array, Format ,400 65,900 52,300 46,600 TaqMan Array, Format ,400 65,900 52,300 46,600 TaqMan Array, Format ,000 70,300 54,500 48,800 TaqMan Array, Format ,000 70,300 54,500 48,800 TaqMan Array, Format ,300 72,600 56,800 49,900 TaqMan Array, Format 96a ,500 73,900 63,600 52,300 TaqMan Array, Format 96b ,500 73,900 63,600 52,300 TaqMan Array, Format ,200 81,700 70,300 56,800 TaqMan Array, Format ,900 95,300 74,900 61,300 * TaqMan Gene Expression Assays, Inventoried 24 TaqMan Gene Expression Assays, Made-to-Order

29 TaqMan TaqMan Array Endogenous Control Plate Applied Biosystems TaqMan Array Endogenous Control Plate TaqMan TaqMan Array Human Endogenous ,700 Controls, 96-well Plate TaqMan Array Human Endogenous ,700 Controls, Fast 96-Well Plate TaqMan Array Mouse Endogenous ,700 Controls, 96-well Plate TaqMan Array Mouse Endogenous ,700 Controls, Fast 96-Well Plate TaqMan Array Rat Endogenous ,700 Controls, 96-well Plate TaqMan Array Rat Endogenous Controls, Fast 96-Well Plate , TaqMan Array Human Endogenous Control Plate Assay ID Gene Symbol TaqMan Human Endogenous Control Array Applied Biosystems TaqMan Human Endogenous Control Array TaqMan Gene Signature Arrays Gene Expression Assays TaqMan Human Endogenous Control Array 7900HT ViiA 7 QuantStudio TaqMan Array TaqMan Human Endogenous ,100 Control Array 29

30 TaqMan TaqMan Gene Expression Assays Endogenous Control Applied Biosystems TaqMan Gene Expression Assays TaqMan Endogenous Controls Applied Biosystems TaqMan Endogenous Controls VIC FAM TaqMan Gene Expression 50 µl ,500 Assays, Inventoried 20 µl 250 TaqMan Endogenous Control 1,000 ** 111,300 VIC/MGB * 50 µl ** Human Gene Symbol RefSeq Gene Name TaqMan Gene Expression Assays Assays ID TaqMan Endogenous Controls *VIC/MGB FAM/MGB 18S X Eukaryotic 18S rrna Hs _s E E ACTB NM_ actin, beta Hs _m E E GAPDH NM_ glyceraldehyde-3-phosphate dehydrogenase Hs _m E E RPLP0 NM_ ribosomal protein, large, P0 Hs _m E PPIA NM_ peptidylprolyl isomerase A (cyclophilin A) Hs _m E PGK1 NM_ phosphoglycerate kinase 1 Hs _m E B2M NM_ beta-2-microglobulin Hs _m E GUSB NM_ glucuronidase, beta Hs _m E HPRT1 NM_ hypoxanthine phosphoribosyltransferase 1 (Lesch-Nyhan syndrome) Hs _m E TBP M TATA box binding protein Hs _m E TFRC NM_ transferrin receptor (p90, CD71) Hs _m E IPO8 NM_ importin 8 Hs _m1 YWHAZ NM_ tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein, zeta polypeptide Hs _m1 UBC NM_ ubiquitin C Hs _m1 POLR2A NM_ polymerase (RNA) II (DNA directed) polypeptide A, 220kDa Hs _m1 HMBS NM_ hydroxymethylbilane synthase Hs _m1 * VIC TaqMan Fast Universal Master Mix /

31 Mouse Gene Symbol RefSeq Gene Name TaqMan Gene Expression Assays Assays ID TaqMan Endogenous Controls *VIC/MGB FAM/MGB 18S X Eukaryotic 18S rrna Hs _s E E ACTB NM_ actin, beta, cytoplasmic Mm _s E E Gapdh NM_ glyceraldehyde-3-phosphate dehydrogenase Mm _g E E Rplp2 (AK ) ribosomal protein, large P2 Mm _s1 Ppia NM_ peptidylprolyl isomerase A Mm _g1 Pgk1 NM_ phosphoglycerate kinase 1 Mm _m1 B2m NM_ beta-2 microglobulin Mm _m1 Gusb NM_ glucuronidase, beta Mm _m1 Hprt1 NM_ hypoxanthine guanine phosphoribosyl transferase 1 Mm _m1 Tbp NM_ TATA box binding protein Mm _m1 Tfrc NM_ transferrin receptor Mm _m1 Ipo8 NM_ importin 8 Mm _m1 Ywhaz NM_ tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein, zeta polypeptide Mm _g1 Ubc ubiquitin C Mm _m1 Polr2a NM_ polymerase (RNA) II (DNA directed) polypeptide A Mm _m1 Hmbs NM_ hydroxymethylbilane synthase Mm _g1 Rat Gene Symbol RefSeq Gene Name TaqMan Gene Expression Assays Assays ID TaqMan Endogenous Controls *VIC/MGB FAM/MGB 18S X Eukaryotic 18S rrna Hs _s E E Actb NM_ actin, beta Rn _m E E Gapdh NM_ glyceraldehyde-3-phosphate dehydrogenase Rn _s E Rplp2 NM_ ribosomal protein, large P2 Rn _g1 Ppia NM_ peptidylprolyl isomerase A Rn _m1 Pgk1 NM_ phosphoglycerate kinase 1 Rn _g1 B2m NM_ beta-2 microglobulin Rn _m1 Gusb NM_ glucuronidase, beta Rn _m1 Hprt NM_ hypoxanthine guanine phosphoribosyl transferase Rn _m1 Tbp NM_ TATA box binding protein Rn _m1 Tfrc M transferrin receptor Rn _m1 Hmbs NM_ hydroxymethylbilane synthase Rn _m1 Ywhaz NM_ tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein, zeta polypeptide Rn _m1 Ubc NM_ ubiquitin C Rn _g1 Ppib NM_ peptidylprolyl isomerase B Rn _m1 Arbp NM_ acidic ribosomal phosphoprotein P0 Rn _g1 * VIC TaqMan Fast Universal Master Mix /

32 RNAi RNAi RNA RNAi RNA 2 RNA dsrna dsrna mrna RNAi RNAi RNAi RNA dsrna RNAi sirna Dicer dsrna sirna small interfering RNA sirna RISC RNA-induced silencing complexes sirna RISC mrna RISC mrna 30 dsrna sirna Dicer sirna shrna short hairpin RNA DNA RNAi 1. sirna RNAi 2 sirna sirna sirna sirna sirna sirna sirna sirna sirna sirna mrna sirna PCR TaqMan mrna TaqMan TaqMan RNAi 32

33 sirna Silencer Select sirna Invitrogen Silencer Select sirna LNA sirna sirna 1/10 1/100 5nM LNA 21. Silencer Select sirna sirna 10 sirna nM sirna HeLa sirna triplicate 48 mrna sirna mrna % sirna 5nM Silencer Select Pre-designed sirna Silencer Select Pre-designed sirna 2 70% RNA Silencer Select Validated sirna Silencer Select Validated sirna 80% mrna Silencer Select Pre-designed sirna % RNA 7. Silencer Select sirna Silencer Select Pre-designed Silencer Select Validated Custom Design Select sirna Custom Select sirna Silencer Select Negative Control #1 sirna Silencer Select Negative Control #2 sirna Silencer Select GAPDH Positive Control sirna Silencer Select Pre-designed sirna 98 Silencer Select Validated sirna RNA 80 RNA Silencer Select sirna sirna Select sirna 21 sirna Silencer Select sirna Silencer Select sirna sirna 2 Silencer Select GAPDH sirna GAPDH GAPDH sirna 22. / RNAi 33

34 sirna Silencer Select Pre-designed sirna Silencer Select Pre-designed sirna, inventoried, standard purity Silencer Select Pre-designed sirna, non-inventoried, standard purity Silencer Select Pre-designed sirna, HPLC purified *1 nmol 4 Silencer Select Validated sirna 1 nmol ,700 5 nmol , nmol , nmol ,200 5 nmol , nmol , nmol , nmol , nmol ,700 Silencer Select Validated sirna, standard purity Silencer Select Validated sirna, HPLC purified *1 nmol 4 1 nmol ,800 5 nmol , nmol , nmol , nmol , nmol ,300 Silencer Select Negative Control sirna Silencer Select Negative Control #1 sirna Silencer Select Negative Control #2 sirna 5 nmol , nmol , nmol ,000 5 nmol , nmol ,500 Silencer Select GAPDH Positive Control sirna Silencer Select GAPDH sirna (Hs, Mm, Rn) Custom Select sirna 5 nmol , nmol , nmol ,000 Custom Select sirna, standard purity Custom Select sirna, HPLC purified 5 nmol , nmol , nmol , nmol , nmol ,800 sirna sirna 3 sirna sirna Silencer Select Human Kinase sirna Library V4 2, Silencer Select Human Phosphatase sirna Library V4 Silencer Select Human 1, GPCR sirna Library V4 Silencer Select Human Nuclear Hormone Receptor sirna Library V4 Silencer Select Human Ion 1, Channel sirna Library V4 Silencer Select Human Protease sirna Library V4 1, Silencer Select Human Druggable 27,096 9, Genome sirna Library V4-96-well Silencer Select Human Druggable 4,149 1, Genome V4 sirna Ext Set - 96-well Silencer Select Human Extended 31,245 10, Druggable Genome sirna Library V4-96-well Silencer Select Human Druggable 27,096 9, Genome sirna Library V4-384-well Silencer Select Human Druggable 4,149 1, Genome V4 sirna Ext Set well Silencer Select Human Genome 33,510 11, sirna V4 Extension Set well Silencer Select Human Genome sirna Library V4-384-well 64,755 21, nmol 0.5 nmol sirna sirna 36 sirna sirna sirna nmol, 0.25 nmol, 1 nmol, 2 nmol sirna Inventoried sirna non- Inventoried sirna 34

35 Ambion In Vivo sirna Invitrogen Ambion In Vivo sirna in vivo RNAi sirna Silencer Select Silencer Select 33 in vivo RNAi LNA 90% RNAi sirna remaining intact (%) Unmodified sirna: time (h) t 1/2 5 min Ambion In Vivo sirna: t 1/2 24 h Ambion In Vivo Pre-designed sirna, Standard Purity Ambion In Vivo Pre-designed sirna,hplc Ambion In Vivo Pre-designed sirna, HPLC In Vivo Ready Ambion In Vivo Custom sirna Standard Purity Ambion In Vivo Custom sirna, HPLC Ambion In Vivo Custom sirna, HPLC In Vivo Ready Ambion In Vivo Custom Designed Standard Purity Ambion In Vivo GAPDH Positive Control sirna Ambion In Vivo GAPDH Positive Control sirna, HPLC In Vivo Ready Ambion In Vivo Factor VII sirna, HPLC In Vivo Ready, 50 nmol Ambion In Vivo Factor VII Positive Control sirna, HPLC In Vivo Ready, 250 nmol Ambion In Vivo Negative Control #1 sirna, 5 nmol Ambion In Vivo Negative Control #1 sirna, HPLC In Vivo Ready, 50 nmol Ambion In Vivo Negative Control #1 sirna, HPLC In Vivo Ready, 250 nmol 5 nmol , nmol ,000 1 µmol , nmol ,000 1 µmol ,000 5 nmol , nmol ,100 1 µmol , nmol ,000 1 µmol ,000 5 nmol ,300 5 nmol , nmol , nmol , nmol , nmol ,000 5 nmol , nmol , nmol , sirna Ambion In Vivo sirna 8 sirna 90% sirna HPLC TaqMan sirna Assays Applied Biosystems TaqMan sirna Assays Silencer Select sirna in vivo sirna TaqMan Assays sirna shrna TaqMan sirna Assays, ,900 Made-to-Order, S TaqMan sirna Assays, ,500 Made-to-Order, M TaqMan sirna Assays, 2, ,600 Made-to-Order, L *20 µl 35

36 RNA sirna mirna Lipofectamine RNAiMAX Invitrogen Lipofectamine RNAiMAX sirna mirna Silencer Select sirna 33 mirvana mirna mimics & inhibitors 43 sirna Invivofectamine 3.0 Reagent Invitrogen Invivofectamine 3.0 Reagent in vivornai sirna 85 sirna 85% sirna 90% p53 発現レベル細胞生存率 (%) p53 発現レベル 細胞生存率 (%) 24. Lipofectamine RNAiMAX Lipofectamine RNAiMAX 試薬量 µl RNAiMAX p Invivofectamine 3.0 Reagent Factor VII PPIB sirnai mrna 85 p53, relative to nontransfected nm + Lipofectamine RNAiMAX Scrambled control 10 nm + B 社製品 10 nm + T 社製品 p53 sirna 10 nm + Q 社製品 1 nm + Lipofectamine RNAiMAX 1 nm + B 社製品 1 nm + T 社製品 1 nm + Q 社製品 Nontransfected 25. Lipofectamine RNAiMAX 27. sirna Invivofectamine 3.0 Reagent FVII sirna mg/kg FVII sirna Invivofectamine 3.0 Reagent ED mg/kg mg/kg Lipofectamine RNAiMAX 0.1 ml , ml , ml , ml , ml ,300 Invivofectamine 3.0 Reagent 10 IVF , IVF ,000 36

37 microrna microrna microrna mirna pol precursor mirna mirna 21-23nt mature mirna mirna non-coding RNA mirna mirna mrna 3'UTR mrna mirna Human Precursor mirna Mature mirna 28. mirna (Mature mirna) mirna mirna mirna mirbase mirna 8. mirna mirna mirna mirna mirvana mirna Isolation Kit mirvana PARIS Kit TaqMan mirna ABC Purlfication Kit TaqMan MicroRNA Cells-to-CT Kit TaqMan Array MicroRNA Cards TaqMan Universal PCR Master Mix II Megaplex Primer Pools TaqMan MicroRNA RT Kit TaqMan MicroRNA Assays TaqMan Universal PCR Master Mix II TaqMan MicroRNA RT Kit mirvana mirna mimic mirvana mirna inhibitor Lipofectamine RNAiMAX TaqMan Gene Expression Cells-to-CT Kit TaqMan Gene Expression Assays 37

38 microrna mirvana mirna Isolation Kit RNA RNA microrna mirna small RNA Invitrogen mirvana mirna Isolation Kit mirna RNA mirna Total RNA 250mg Total RNA 30 mirvana mirna Isolation Kit 40 AM ,000 mirvana mirna Isolation Kit without Phenol mirvana PARIS Kit 40 AM ,800 small RNA RNA 2 1 Small RNA RNA mirna 30 small RNA RNA mrna mirna sirna mg 200 small RNA TaqMan mirna ABC Anti-miRNA Bead Capture Purification Kit mirna 75 Human Panel A B 754 mirna mirna Anti-miRNA oligos Dynabeads mirna mirna 10µL 50µL RT-qPCR TRIzol 75 mirna Avg. C t cel-mir-39 expression profile serum sample (50 µl) hsa-mir-16 hsa-let-7a Assay ID 30. TaqMan mirna ABC Purification Ki 50µL hsa-mir-150 hsa-let-7c hsa-mir-135b RNA mirvana PARIS Kit 40 AM , ABC Purification Kit TaqMan mirna ABC Purification ,500 Kit-Human Panel A TaqMan mirna ABC Purification Kit-Human Panel B ,500 38

39 microrna TaqMan MicroRNA RT Kit Applied Biosystems TaqMan MicroRNA RT Kit TaqMan MicroRNA Assays 40 TaqMan MicroRNA Assays RT TaqMan MicroRNA RT Kit ,300 Megaplex Primer Pools 1, ,800 Applied Biosystems Megaplex RT Primers stem-looped Sanger mirbase mirna mirna RNA Megaplex Primer Pools TaqMan MicroRNA Assays TaqMan MicroRNA Arrays Megaplex PreAmp Primers 1ng RNA Megaplex RT Primers Pool Rodent Megaplex RT Primers, Rodent Pool A ,600 Megaplex RT Primers, Rodent Pool B v ,600 Megaplex RT Primers, Rodent Pool Set v3.0 RT Primers Pools A&B & ,400 Megaplex PreAmp Primers Pool Rodent Megaplex PreAmp Primers, ,600 Rodent Pool A PreAmp Megaplex PreAmp Primers, ,600 Rodent Pool B v3.0 PreAmp Megaplex PreAmp Primers, Rodent Pool Set v3.0 PreAmp Primers Pools A&B & ,400 Megaplex RT and PreAmp Primers Package Rodent Megaplex Primer Pools, ,300 Rodent Pools A RT Primers PreAmp Primers Megaplex Primer Pools, ,300 Rodent Pools B v3.0 RT Primers PreAmp Primers Megaplex Primer Pools, Rodent Pools Set v3.0 RT Primers Pools A&B PreAmp Primers Pools A&B ,500 Megaplex RT Primers Pool Human Megaplex RT Primers, Human Pool A ,600 Megaplex RT Primers, Human Pool B v ,600 Megaplex RT Primers, Human Pool Set v3.0 RT Primers Pools A&B & ,400 Megaplex PreAmp Primers Pool Human Megaplex PreAmp Primers, ,600 Human Pool A PreAmp Megaplex PreAmp Primers, ,600 Human Pool B v3.0 PreAmp Megaplex PreAmp Primers, Human Pool Set v3.0 PreAmp Primers Pools A&B & ,400 Megaplex RT and PreAmp Primers Package Human Megaplex Primer Pools, ,300 Human Pools A RT Primers PreAmp Primers Megaplex Primer Pools, ,300 Human Pools B v3.0 RT Primers PreAmp Primers Megaplex Primer Pools, Human Pools Set v3.0 RT Primers Pools A&B PreAmp Primers Pools A&B ,500 TaqMan MicroRNA Arrays A B Megaplex Primer Pools 39

40 microrna TaqMan MicroRNA Assays Applied Biosystems TaqMan MicroRNA Assays PCR microrna mirna TaqMan MicroRNA Assays mirna mirna mirna mirna Assays Synthetic mirna Target Relative Detection (%) No. of base differences TaqMan mirna mirna PCR ng RNA 7 TaqMan MicroRNA Assays TaqMan MicroRNA Assays mirna loop mirna RT mirna mirna 5' TaqMan PCR mirna Mature mirna Looped RT primer TaqMan MicroRNA Assays 1 let-7a let-7a let-7c 3.7 TaqMan MicroRNA Assays, Inventoried* mirna RT primer 5X reaction mix (50 / 15 µl ) mirna TaqMan Assay 20X reaction mix (150 / 20 µl ) ,500 TaqMan MicroRNA Assays, Made-to-Order, Small mirna RT primer 5X reaction mix (50 / 15 µl ) mirna TaqMan Assay 20X reaction mix (150 / 20 µl ) TaqMan MicroRNA Assays, Made-to-Order, Medium TaqMan MicroRNA Assays, Made-to-Order, Large ,900 2, , , ,600 * Arabidopsis thaliana Caenorhabditis elegans Drosophila melanogaster Made-to-Order Sanger Release 16 mirna Custom TaqMan Small RNA Assays Forward primer TaqMan probe Reverse primer 32. TaqMan MicroRNA Assays TaqMan MicroRNA Assays mirna mirnas Custom TaqMan Small RNA Assays, ,200 Custom TaqMan Small RNA Assays, Custom TaqMan Small RNA Assays, *20 µl ,900 2, , ,300 40

41 TaqMan Array MicroRNA Card Applied Biosystems TaqMan Array MicroRNA Card TaqMan 384 TaqMan mirna 1 Megaplex RT Primers 39 TaqMan MicroRNA Array v2.0 / v3.0 Megaplex PreAmp Primers TaqMan MicroRNA Arrays v2.0 / v3.0 Megaplex Primer Pools Sanger mirbase 5 TaqMan Array Applied Biosystems 7900HT ViiA 7 QuantStudio TaqMan Array Human TaqMan Array Human MicroRNA A Cards v2.0 TaqMan Array Human MicroRNA A+B Cards Set v3.0 A array ( ) + B Array ( ) TaqMan Array Human MicroRNA B Cards v3.0 Rodent , , ,000 TaqMan Array Rodent MicroRNA A Cards v2.0 TaqMan Array Rodent MicroRNA A+B Cards Set v3.0 A array ( ) + B Array ( ) TaqMan Array Rodent MicroRNA B Cards v , , ,000 mature mirna mirvana mirna Isolation Kit TaqMan mirna ABC Purification Kit Megaplex RT Primers & TaqMan MicroRNA Reverse Transcription Kit Megaplex PreAmp Primers & TaqMan PreAmp Master Mix Diluted RT product & TaqMan Universal PCR Master Mix II Applied Biosystems QuantStudio 12K Flex ViiA 7 / 7900 HT Fast PCR RT-PCR Expression Suite Software 34. mirna 41

42 Non-coding RNA TaqMan Pri-miRNA Assays mirna TaqMan Pri-miRNA Assays Applied Biosystems TaqMan Pri-miRNA Assays microrna mirna primary microrna Pri-miRNA microrna Pri-miRNA 1 microrna Drosha mirna mirna stem-loop Dicer RNA Induced Silencing Complex RISC mirna 35. microrna 37. hsa-mir-26 mir-26a-1 mir-26a-2 Pri-miRNA Stemloop Pri-miR-26a-2 TaqMan Pri-miRNA Assay hsa-mir-26a-2 TaqMan Non-coding RNA Assays TaqMan Non-coding RNA Assays Applied Biosystems TaqMan Non-coding RNA Assays non-cording TaqMan Gene Expression Assays TaqMan Non-coding RNA TaqMan Non-coding RNA Assays TaqMan PreAmp Master Mix Pre-Amplification 36. TaqMan Pri-miRNA Assays Sanger mirbase Stem-loop 500 Pri-miRNA Sanger mirbase sequence repository stem-loop mirna TaqMan Pri-miRNA Assay TaqMan MicroRNA Assay mirna TaqMan Non-coding RNA Assays, S ,900 TaqMan Non-coding RNA Assays, M ,500 TaqMan Non-coding RNA Assays, L 2, ,600 *20 µl TaqMan Pri-miRNA Assays, S ,900 TaqMan Pri-miRNA Assays, M ,500 TaqMan Pri-miRNA Assays, L 2, ,600 *20 µl 42

43 microrna mirvana mirna mimic & inhibitors mirna mirna mirvana mirna inhibitor Invitrogen mirvana mirna inhibitor mirna mirna mirna 1 mirbase in vivo in vivo ready mirvana mirna mimic Invitrogen mirvana mirna mimic mirna mirna mirna mirna 2 mirna mirvana mirna mimic 5 nmol ,900 mirvana mirna mimic, in vivo Ready Customer-Defined mirvana mirna mimic Customer-Defined mirvana mirna mimic, in vivo Ready mirvana mirna mimic Negative Control #1 mirvana mirna mimic Negative Control #1, in vivo Ready mirvana mirna mimic mir-1 Positive Control mirvana mirna mimic mir-1 Positive Control, in vivo Ready mirvana mirna mimic mirbase v nmol , nmol ,500 5 nmol , nmol , nmol ,500 5 nmol , nmol , nmol , nmol ,000 5 nmol , nmol , nmol , nmol , nmol each 1 nmol each mirvana mirna inhibitor 5 nmol , nmol ,900 mirvana mirna inhibitor, 250 nmol ,500 in vivo Ready Customer-Defined mirvana 5 nmol ,900 mirna inhibitor 20 nmol ,000 Customer-Defined mirvana 250 nmol ,500 mirna inhibitor, in vivo ready mirvana mirna inhibitor 5 nmol ,900 Negative Control #1 20 nmol ,700 mirvana mirna inhibitor 50 nmol ,500 Negative Control #1, in vivo Ready 250 nmol ,000 mirvana mirna inhibitor let-7 5 nmol ,900 Positive Control 20 nmol ,700 mirvana mirna inhibitor mirbase v Pre-miR mirna Precursor Molecule Pre-miR mirna Precursor 1 AM ,700 Starter Kit Pre-miR mirna Precursors 5 nmol AM ,800 Molecule 20 nmol AM , nmol Customer-Defined Pre-miR 20 nmol AM ,200 mirna Precursor 4 5 nmol Pre-miR Negative Control #1 5 nmol AM ,800 Pre-miR Negative Control #2 5 nmol AM ,800 Cy3 Pre-miR Negative Control 5 nmol AM ,100 FAM Pre-miR Negative Control 5 nmol AM ,900 Pre-miR hsa-mir-1 mirna Precursor 5 nmol AM ,800 Anti-miR mirna Inhibitor Anti-miR mirna Inhibitor 5 nmol AM , nmol 4 5 nmol AM ,200 Customer Defined Anti-miR mirna Inhibitor 0.25 nmol each 1 nmol each 20 nmol 4 5 nmol AM ,200 Anti-miR Negative Control #1 5 nmol AM ,800 Cy3 Anti-miR Negative Control 5 nmol AM ,000 FAM Anti-miR Negative Control 5 nmol AM ,900 mirna 36 Lipofectamine RNAiMAX Invivofectamine

44 SNP TaqMan SNP 2 TaqMan TaqMan 5' VIC dye FAM dye 3' Q PCR DNA VIC dye FAM dye TaqMan SNP Genotyping Assay 100 DNA 38. TaqMan SNP 39. Allele1 TaqMan VIC Allele2 TaqMan FAM VIC FAM Allele1 Allele2 Allele1 2 44

45 SNP TaqMan Pre-Designed SNP Genotyping Assays Applied Biosystems TaqMan SNP Genotyping Assays SNP SNP SNP ID SNP csnp 7 SNP 16 SNP 350 SNP 2600 Human TaqMan Pre-Designed SNP Genotyping Assays, Human, non-inventried Small scale 300 1, ,600 TaqMan Pre-Designed SNP Genotyping Assays, Human, non-inventried Medium scale TaqMan Pre-Designed SNP Genotyping Assays, Human, non-inventried Large scale * 25 µl 5 µl non-human 1,000 5,000 2,400 12, , ,900 TaqMan Pre-Designed SNP Genotyping Assays Non-Human non-inventoried Small scale 300 1, ,600 TaqMan Pre-Designed SNP Genotyping Assays Non-Human non-inventoried Medium scale TaqMan Pre-Designed SNP Genotyping Assays Non-Human non-inventoried Large scale * 25 µl 5 µl 1,000 5,000 2,400 12, , ,900 Custom TaqMan SNP Genotyping Assays Applied Biosystems Custom TaqMan SNP Genotyping Assays SNP SNP SNP SNP SNP 6 MNP SNP Human Custom TaqMan SNP Genotyping ,900 Assays, S-scale, Human 1,500 Custom TaqMan SNP Genotyping Assays, M-scale, Human Custom TaqMan SNP Genotyping Assays, L-scale, Human * 25 µl 5 µl non-human 1,000 5,000 2,400 12, , ,700 Custom TaqMan SNP Genotyping ,900 Assays, S-scale, non-human 1,500 Custom TaqMan SNP Genotyping Assays, M-scale, non-human Custom TaqMan SNP Genotyping Assays, L-scale, non-human * 25 µl 5 µl 1,000 5,000 2,400 12, , ,700 TaqMan Drug Metabolism Genotyping Assays 220 2,000 TaqMan Drug Metabolism ,000 Genotyping Assays 750 * 25 µl 5 µl 45

46 SNP SNP TaqMan Genotyping Master Mix Applied Biosystems TaqMan Genotyping Master Mix PCR GC AT TaqMan Universal Master Mix GC AT 72 Fast SNP TaqMan GTXpress Master Mix Applied Biosystems TaqMan GTXpress Master Mix Fast PCR SNP SNP TaqMan PCR 50 PCR 50 Tracking Dye 9. PCR 40. TaqMan Genotyping Master Mix GC-rich TaqMan Genotyping Master Mix 94 gdna TaqMan SNP Assay C _10 PCR GeneAmp PCR System HT PCR GC-rich Catjespin D rs12214 dbsnp TaqMan Genotyping Master Mix TaqMan Genotyping Master Mix, ,800 1 ml 400 TaqMan Genotyping Master Mix, 10 ml TaqMan Genotyping Master Mix, 10 ml 2 TaqMan Genotyping Master Mix, 50 ml TaqMan Genotyping Master Mix, 50 ml 2 TaqMan Genotyping Master Mix TaqMan Genotyping Master Mix 800 4,000 1,600 8,000 4,000 20,000 8,000 40, , , , ,089, TaqMan GTXpress Master Mix TaqMan GTXpress Master Mix, 1 ml ,800 TaqMan GTXpress Master Mix, 10 ml TaqMan GTXpress Master Mix, 50 ml TaqMan GTXpress Master Mix, 100 ml 2 50 ml *PCR 96 Standard Block,0.2 µl 25 µl 96 Fast Block,0.1 µl 10 µl µl 800 2,000 4,000 4,000 10,000 20,000 8,000 20,000 40, , , ,089,800 * 25 µl 5 µl 46

47 SNP TaqMan Sample-to-SNP Kit Applied Biosystems TaqMan Sample-to-SNP Kit DNA TaqMan SNP 1 DNA Fast PCR TaqMan GTXpress Master Mix DNA 5 PCR 50 SNP TaqMan Sample-to-SNP Kit / FTA / / / Buccal swab / Rat Mouse tail / Leaf punch / / FFPE DNA 95 3 DNA PCR A. Buccal swab, B. Mouse tail, C. Corn leaf, D. FFPE Fast PCR TaqMan GTXpress Master Mix PCR 50 TaqMan Sample-to-SNP Kit 5 ml sample prep, 1 ml PCR TaqMan Sample-to-SNP Kit 20 ml sample prep, 10 ml PCR TaqMan Sample-to-SNP Kit 20 ml sample prep, 50 ml PCR TaqMan Sample-to-SNP Kit 200 ml sample prep, 10 ml PCR * PCR 25 µl DNA 100 PCR 80 DNA 400 PCR 800 DNA 400 PCR 4,000 DNA 4,000 PCR , , , , TaqMan Sample-to-SNP Kit SNP SNP / 47

48 TaqMan Mutation Detection Assays Applied Biosystems TaqMan Mutation Detection Assays Competitive Allele-Specific TaqMan PCR castpcr MGB TaqMan Mutation Detection Assay 11. アッセイのタイプ 説明 模式図 Reference Assay Mutant Assay Mutant Allele Assay 特定の変異型アレルを検出します (EGFR_exlqdels_mu* は 19 種類の変異を検出 ) アレル特異的プライマーが変異型アレルを検出します MGB ブロッカーオリゴスクレオチドが野生型アレルを抑えます ASP = 対立遺伝子特異的プライマー ASB = 対立遺伝子特異的ブロッカー (MGB) LST = 遺伝子特異的 TaqMan プローブ LSP = 遺伝子特異的プライマー EGFR exon19 deletion Wild type Assay Mutant Assay EGFR Reference Assay EGFR Exon19 Deletion Assay Wild type Allele Assay 野生型アレルを検出します アレル特異的プライマーが野生型アレルを検出します MGB ブロッカーオリゴヌクレオチドが変異型アレルを抑えます ASP = 対立遺伝子特異的プライマー ASB = 対立遺伝子特異的ブロッカー (MGB) LST = 遺伝子特異的 TaqMan プローブ LSP = 遺伝子特異的プライマー Mutation Detector TaqMan Mutation Detection Assay Gene Reference Assay 変異を含む遺伝子を検出します 遺伝子特異的なプライマーペアで遺伝子内の変異のない領域を増幅します FP = フォワードプライマー RP = リバースプライマー LST = 遺伝子特異的 TaqMan プローブ ASP: LST: TaqMan LSP: MGB: ASB: MGB FP: RP: TaqMan Mutation Detection Assays ** 150 µl ,800 TaqMan EGFR Exon 19 Deletions Assay 1 EGFR 19 1 ** 150 µl ,800 TaqMan Mutation Detection Reference Assays 46 ** 150 µl ,200 TaqMan Mutation Detection IPC Reagent Kit 1 kit ,800 *20 µl ** TaqMan Mutation Detection Assays 48

49 TaqMan Copy Number Assays Copy Number Variations, CNVs CNVs CNVs Applied Biosystems TaqMan Copy Number Assays CNVs 160 TaqMan CNVs TaqMan Copy Number Assays TaqMan Copy Number Reference Assays PCR 3 4 PCR DNA Copy Number 4 TaqMan Copy Number 360 assays ,700 Assays, Small 720 assays TaqMan Copy Number Assays, Medium TaqMan Copy Number Assays, Large 750 assays 1,500 assays 2,900 assays 5,800 assays , ,600 * 20 µl µl 384 Custom TaqMan Copy Number Assays Applied Biosystems Custom TaqMan Copy Number Assays TaqMan Copy Number Assays CNVs GeneAssist Copy Number Assay Tool CNVs Custom TaqMan Copy Number 360 assays ,200 Assays, Small 720 assays Custom TaqMan Copy Number Assays, Medium Custom TaqMan Copy Number Assays, Large 750 assays 1,500 assays 2,900 assays 5,800 assays , ,300 * 20 µl µl 384 CopyCaller TaqMan Copy Number Assays Applied Biosystems CopyCaller TaqMan Copy Number Reference Assays Applied Biosystems TaqMan Copy Number Reference Assays TaqMan Copy Number Assays Custom TaqMan Copy Number Assays RNase P Ribonuclease P RNA component H1 (H1RNA) gene (RPPH1) on chromosome 14, cytoband 14q11.2 TERT RNase P telomerase reverse transriptase (TERT) gene located on chromosome 5, cytoband 5p15.33 Human TaqMan Copy Number 750 assays ,500 Reference Assay, RNase P 1,500 assays 3,000 assays 6,000 assays ,900 TaqMan Copy Number Reference Assay, TERT 750 assays 1,500 assays 3,000 assays 6,000 assays , ,900 Mouse TaqMan Copy Number 750 assays ,500 Reference Assay, TFRC 1,500 assays 3,000 assays 6,000 assays ,900 TaqMan Copy Number Reference Assay, TERT 750 assays 1,500 assays 3,000 assays 6,000 assays , ,900 * 20 µl µl 384 Custom Plus TaqMan Copy Number Assays Applied Biosystems Custom Plus TaqMan Copy Number Assays Custom TaqMan Copy Number Assays SNP TaqMan TaqMan Copy Number Assays Custom Plus TaqMan 360 assays ,200 Copy Number Assays, Small 720 assays Custom Plus TaqMan Copy Number Assays, Medium Custom Plus TaqMan Copy Number Assays, Large 750 assays 1,500 assays 2,900 assays 5,800 assays , ,300 * 20 µl µl

50 High Resolution Melt HRM High Resolution Melting HRM PCR PCR PCR Applied Biosystems MeltDoctor HRM MeltDoctor HRM ステップ 1: 装置のキャリブレーション * ステップ 2: PCR の準備 ステップ 3: PCR ステップ 4: HRM 解析 ステップ 5: データ解析 HRM の蛍光色素と プライマーの設計を行い PCR 増幅を行います リアルタイム PCR で 融解曲線のデータを バックグラウンドの 必要に応じて PCR 条件の ステップ 3 ~ 5 はリアルタイム 融解曲線を描きます 解析します キャリブレーションを 最適化を行います PCR を使った場合の一連の 行います 操作となります *Only required once every six months 43. HRM HRM PCR MeltDoctor HRM High Resolution Melting HRM PCR MeltDoctor HRM SYTO 9 DNA Tm PCR dutp dntp PCR HRM 50

51 MeltDoctor HRM HRM AmpliTaq Gold 360 DNA Polymerase PCR MeltDoctor HRM 2 SYTO 9 MeltDoctor HRM MeltDoctor HRM HRM PCR MeltDoctor HRM units ,400 AmpliTaq Gold 360 DNA Polymerase GeneAmp dntp Blend MeltDoctor HRM Dye MeltDoctor HRM ,000 5 ml MeltDoctor HRM 2, , ml MeltDoctor HRM 5, , ml MeltDoctor HRM 5, , ml *20 µl MeltDoctor HRM Applied Biosystems MeltDoctor HRM 3 AA, AB, BB 3 DNA MeltDoctor HRM ,000 MeltDoctor HRM Applied Biosystems MeltDoctor HRM 96 Fast MeltDoctor HRM DNA 44. HRM 3 HeLa, Raji, and Jurkat HRM p bp PCR 3 HRM GC, GG, CC HRM MeltDoctor HRM Applied Biosystems 7500 Fast PCR Applied Biosystems HRM Software v2.0 HRM PCR BigDye Terminator v1.1 Cycle Sequencing Kit Applied Biosystems 3130 MeltDoctor HRM , Fast MeltDoctor HRM , MeltDoctor HRM 20 1 ml ,000 HRM HRM HRM PCR 51

52 TaqMan Protein Assays 市販または自家調製抗体 SA- オリゴ A 3 末端オリゴ アッセイプローブ A 5 末端オリゴ SA- オリゴ B アッセイプローブ B エピトープ 1 エピトープ 2 Build Your Assay 90 min Prepare Sample 30 min Binding 60 min - TaqMan Protein Assay Open Kit Protein Quant Sample Lysis Kit Protein Expression Sample Lysis Kit Ambion PARIS Kit TaqMan Protein Assay TaqMan Protein Assay Open Kit TaqMan Protein Assay Kit 高濃度中濃度低濃度 コネクター 蛍光強度 バックグラウンド 相対発現量変化 サイクル数 Ligation/ Inactivation 25 min TaqMan Fast 40 min Real-Time PCR Data Analysis DNA TaqMan Protein Expression Core Reagents Kit with Master Mix Universal TaqMan Assay PCR TaqMan PCR mrna mirna ProteinAssist Software v TaqMan Protein Assays 52

53 TaqMan Protein Assays Applied Biosystems TaqMan Protein Assays TaqMan 5 mrna mirna 500 TaqMan Protein Assays Kit hesc Applied Biosystems TaqMan Protein Assays Kit hicam1 hcstb TaqMan Protein Assay Kit hcstb ,600 TaqMan Protein Assay Kit hicam ,600 4 TaqMan Protein Expression Core Reagents Kit with Master Mix Applied Biosystems TaqMan Protein Expression Core Reagents Kit with Master Mix PCR TaqMan Protein Assays Kit TaqMan Protein Assays Open Kit TaqMan Protein Assay Core Reagent Base Kit DNA Ligase, Protease, Ligase PCR / TaqMan Protein Assays Fast Master Mix ph PCR TaqMan Protein Assay Open Kit Applied Biosystems TaqMan Protein Assay Open Kit TaqMan Protein Assay 2 TaqMan PCR mrna mirna TaqMan Protein Assays Open Kit 4, ,400 TaqMan Protein Assays Oligo Probe Kit TaqMan Protein Assays Buffer Kit TaqMan Protein Assays Oligo 4, ,400 Probe Kit 5'Prox-Oligo 200 nm, 50 µl 3'Prox-Oligo 200 nm, 50 µl TaqMan Protein Assays Buffer Kit Antibody Dilution Buffer, 1 ml Assay Probe Strage Buffer, 1 ml Assay Probe Dilution Buffer, 7 ml Lysate Dilution Buffer, 25 ml 4, ,300 TaqMan Protein Expression Core Reagents Kit with Master Mix ,900 TaqMan Protein Expression Core Reagents Base Kit TaqMan Protein Assays Fast Master Mix TaqMan Protein Expression Core Reagents Kit with Master Mix ,000 TaqMan ProteinExpression Core Reagents Base Kit TaqMan Protein Assays Fast Master Mix TaqMan Protein Assays ,500 Core Reagents Base Kit ,700 TaqMan Protein Assays ,700 Fast Master Mix ,600 53

54 Protein Expression Sample Preparation Kit TaqMan Protein Assay Kit TaqMan Protein Assay Open Kit 10,000 50,000 Protein Quant Sample Lysis Kit 50, Protein Expression Sample Preparation Kit Protein Quant Sample Lysis Kit 10,000 50,000 50,000 Lysis Yes No TaqMan Protein Assays 500 cells/µl 2,500 cells/µl TaqMan Protein Assay 1:2 1:10 Protein Expression Lysate Control Kits Applied Biosystems TaqMan Protein Expression Lysate Control Kits Protein Expression Lysate Control Kit Raji hcstb hicam1 Protein Expression Lysate Control Kit ,100 Raji ProteinAssist Applied Biosystems TaqMan ProteinAssist ProteinAssist Protein Expression Sample Prep Kit 1, ,800 cell counting Protein Quant Sample Lysis Kit cell counting or total protein quant 25 ml ,800 54

55 PCR Protein Thermal Shift Applied Biosystems Protein Thermal Shift PCR PCR 1 10 * PCR * 1 Protein Thermal Shift , Protein Thermal Shift ,800 Protein Thermal Shift 2, ,400 Dye Kit Protein Thermal Shift Starter Kit 2, , ,000 Protein 80,000 Protein + Ligand d /dt 60,000 40,000 20, ,000 10,000 5, , ~7oC Tm Difference Temperature (oc) StepOne /StepOnePlus 7500 / 7500 Fast QuantStudio 7 / QuantStudio 12K Flex PCR 46. Tm 30 PCR 47. Protein Thermal Shift 55

56 13. ル シ イ 384-Well Clear Optical Reaction Plate ( 製品番号 ) 96-Well Reaction Plate ( 製品番号 N / ) + + Adhesive Film ( 製品番号 / ) Adhesive Film ( 製品番号 / ) QuantStudio 12K Flex リアルタイム PCR システム QuantStudio 12K Flex 96-Well Reaction Plate ( 製品番号 N / ) Optical Tubes ( 製品番号 N ) or and + Optical Caps (Flat Cap) ( 製品番号 ) Optical 8-Tube Strip ( 製品番号 ) + + Tray / Retainer Set ( 製品番号 ) Optical Caps (Flat Cap) ( 製品番号 ) QuantStudio 7 Fast 8-Tube Strip, 0.1mL ( 製品番号 ) + 96-Well Tray for VeriFlex Blocks ( 製品番号 ) + Optical 8-Cap Strip ( 製品番号 ) Fast Reaction Tube with Cap, 0.1mL ( 製品番号 ) + 96-Well Tray for VeriFlex Blocks ( 製品番号 ) QuantStudio 7 リアルタイム PCR システム Fast 96-Well Reaction Plate ( 製品番号 / ) + Adhesive Film ( 製品番号 / ) QuantStudio 3 QuantStudio 5 (96 ウェル ) 96-Well Reaction Plate ( 製品番号 N / ) + Optical 8-Tube Strip ( 製品番号 ) Adhesive Film ( 製品番号 / ) + + Tray / Retainer Set ( 製品番号 ) Optical 8-Cap Strip ( 製品番号 ) QuantStudio 3 / QuantStudio 5 リアルタイム PCR システム QuantStudio 3 QuantStudio 5 (Fast 96 ウェル ) Fast 96-Well Reaction Plate ( 製品番号 / ) + Fast 8-Tube Strip, 0.1mL ( 製品番号 ) Adhesive Film ( 製品番号 / ) Well Tray for VeriFlex Blocks ( 製品番号 ) Optical 8-Cap Strip ( 製品番号 ) QuantStudio 5 (384 ウェル ) 384-Well Clear Optical Reaction Plate ( 製品番号 ) + Adhesive Film ( 製品番号 / ) 384-Well Clear Optical Reaction Plate ( 製品番号 ) + Adhesive Film ( 製品番号 / ) Applied Biosystems 7900HT Fast リアルタイム PCR システム 7900HT 96-Well Reaction Plate ( 製品番号 N / ) 96-Well Reaction Plate ( 製品番号 N / ) + + Adhesive Film ( 製品番号 / ) Optical 8-Cap Strip ( 製品番号 ) + Compression Pad ( 製品番号 ) Fast 96-Well Reaction Plate ( 製品番号 / ) + Adhesive Film ( 製品番号 / ) 7500 Fast Fast 96-Well Reaction Plate ( 製品番号 / ) Fast 8-Tube Strip, 0.1mL ( 製品番号 ) + + Adhesive Film ( 製品番号 / ) Optical 8-Cap Strip ( 製品番号 ) Applied Biosystems 7500 Fast / 7500 リアルタイム PCR システム Well Reaction Plate ( 製品番号 N / ) + Adhesive Film ( 製品番号 / ) 7300 Optical Tubes or Optical 8-Tube Strip Optical 8-Cap Strip 注 :Optical 8-Tube Strip を使用の ( 製品番号 N ) and ( 製品番号 ) + ( 製品番号 ) 場合には縦向きにセット Fast 96-Well Reaction Plate ( 製品番号 / ) + Adhesive Film ( 製品番号 / ) StepOne Plus Fast 8-Tube Strip, 0.1mL ( 製品番号 ) Fast Reaction Tube with Cap, 0.1mL ( 製品番号 ) Well Tray for VeriFlex Blocks ( 製品番号 ) 96-Well Tray for VeriFlex Blocks ( 製品番号 ) + Optical 8-Cap Strip ( 製品番号 ) StepOnePlus / StepOne リアルタイム PCR システム StepOne Fast Optical 48-Well Reaction Plate ( 製品番号 ) Fast 8-Tube Strip, 0.1mL ( 製品番号 ) Well Optical Adhesive Film ( 製品番号 / ) Fast 48-Well Tray ( 製品番号 ) + Optical 8-Cap Strip ( 製品番号 ) Fast Reaction Tube with Cap, 0.1mL ( 製品番号 ) + Fast 48-Well Tray ( 製品番号 ) 56

57 Fast 96-Well 7900HT Fast 96 / 7500 Fast / StepOnePlus ViiA 7 / QuantStudio Fast 96 MicroAmp Fast 96-Well Reaction Plate 0.1 ml MicroAmp Fast Optical 96-Well Reaction Plate with Barcode 0.1 ml 96-Well , , , HT 96 / 7700 / 7500 / 7300 / 7000 / 5700 ViiA 7 / QuantStudio Standard 96 MicroAmp Optical 96-Well Reaction Plate 10 N ,200 MicroAmp Optical 96-Well Reaction Plate ,800 with Barcode MicroAmp Optical 96-Well Reaction Plate with Barcode ,900 MicroAmp Optical 96-Well Reaction Plate with Barcode MicroAmp Optical 96-Well Reaction Plate with Barcode and Optical Caps ,700 MicroAmp Optical 96-Well Reaction Plate with Barcode MicroAmp Optical 8-Cap Strip Well 7900HT 384 / ViiA 7 / QuantStudio 384 MicroAmp Optical 384-Well Reaction ,200 Plate with Barcode MicroAmp Optical 384-Well Reaction ,300 Plate with Barcode MicroAmp Optical 384-Well Reaction Plate with Barcode MicroAmp Optical 384-Well Reaction 1, ,400 Plate with Barcode Optical 384-Well Reaction Plate 1000 Code 128 MicroAmp Optical 384-Well Reaction Plate 1, ,900 Optical 384-Well Reaction Plate 1000 EnduraPlate 96-Well Fast MicroAmp EnduraPlate Optical 96-Well Fast Plate Clear , ,000 Multicolor Clear Yellow Blue Green Red Endura 5, ,000 Yellow ,000 Blue ,000 Green ,000 Red , Well MicroAmp EnduraPlate Optical 96-Well Clear Multicolor Clear Yellow Blue Green Red , ,100 8, ,100 Yellow ,500 Blue ,500 Green ,500 Red , Well MicroAmp EnduraPlate Optical 384-Well Clear Multicolor Clear Yellow Blue Green Red , ,300 9, ,300 Yellow ,000 Blue ,000 Green ,000 Red ,000 57

58 7900HT / 7700 / 7500 / 7500 Fast / 7300 / 7000 / 5700 / StepOnePlus / ViiA 7 / QuantStudio MicroAmp Optical Adhesive Film , ,600 MicroAmp Adhesive Film Applicator , Well Compression Pad 7900HT* / 7700 / 7000 / Optical Adhesive Film+Compression Pad *Fast TaqMan Low Density Array Compression Pad MicroAmp Optical Adhesive Film Kit ,700 MicroAmp Optical Adhesive Film 20 1 MicroAmp Optical Film Compression Pad HT 96 * / 7700 / 7500 / 7500Fast / 7000 / 5700 / StepOnePlus / StepOne / ViiA 7 / QuantStudio *384 TaqMan Array Optical Cap MicroAmp Optical 8-Cap Strip Flat ,900 N / 7500 / 7000 / 5700 / ViiA 7 / QuantStudio Standard 96 MicroAmp Optical 8-Tube Strip 0.2 ml MicroAmp Optical Tube without Cap 0.2 ml ,900 2,000 N ,000 58

59 Fast 7500Fast / StepOnePlus / StepOne / ViiA 7 / QuantStudio Fast 96 MicroAmp Fast 8-Tube Strip 0.1 ml ,400 & 7700 / 7000 / 5700 MicroAmp 96-Well Tray/Retainer Set ,700 Fast StepOnePlus / StepOne MicroAmp Fast Reaction Tube with Cap 0.1 ml 1, ,800 StepOne 48-Well StepOne 48 MicroAmp Fast Optical 48-Well ,500 Reaction Plate MicroAmp Optical 48-Well Adhesive ,100 Film ,700 MicroAmp Fast 48-Well Trays ,500 MicroAmp 48-Well Base Adaptor , *StepOne MicroAmp Splash Free 96-Well Base ,500 MicroAmp 96-Well Support Base * 15,600 StepOnePlus StepOnePlus 96 MicroAmp 96-Well Tray for VeriFlex Blocks ViiA 7 / QuantStudio ,200 ViiA 7 / QuantStudio Standard 96 MicroAmp 96-Well Tray/Retainer Set for Veriti System ,800 Compression Pad 7900HT* / 7700 / 7000 / Optical Adhesive Film Compression Pad *Fast TaqMan Low Density Array Compression Pad MicroAmp Optical Film Compression Pad ,600 ViiA 7 / QuantStudio Fast 96 MicroAmp 96-Well Tray for VeriFlex Blocks ,200 LAMP for GeneAmp 5700 and ABI PRISM 7000 System LAMP for Applied Biosystems 7300/7500 Replacement Lamp for ViiA 7/ QuantiStudio 6 / QuantStudio , , ,700 59