MicroRNA RNA mirna RNA [1*] mirna mirna [2*] mirna RNA mirna mirnasmall RNA mirna mirna mirna 2 *Reference

Transcription

1 microrna microrna having a big impact on biology

2 MicroRNA RNA mirna RNA [1*] mirna mirna [2*] mirna RNA mirna mirnasmall RNA mirna mirna mirna 2 *Reference

3 Contents 4 small-rna 8 microrna 13 mirna 18 microrna 23 microrna 26 3

4 small-rna small RNA 2 10,000 small RNA mirbase [3] smallrna Ion PGM RNA small RNA Ion Ion PGM small RNA Ion Total RNA-Seq v2 1 ng small RNAsmall RNA cdna Ion Total RNA-Seq v2 Ambion US Patent: ' 5' Ion OneTouch 3 Ion PGM 1 Torrent Suite 1 small RNA small RNAcDNA cdna small RNA cdna small RNA small RNA TaqMan 5'nuclease 1 1 ライブラリ作製 2 テンプレート調製 3 シーケンス 4 データ解析 6 時間 5 時間 2 時間 1 時間 Ion Total RNA-Seq Kit v2 Ion OneTouch システム Ion PGM シーケンサ Ion Proton シーケンサ Torrent Server Torrent Suite Software 1 small-rna 4

5 Ion small RNA-Seq small RNA Ion RNA-Seq kit v2 Ion 314 Ion 316 mirna 3 Ion 314 3Ion GeneChip mirna 3 TaqMan Array MicroRNA 2 mirnaion PGM 1miRNA Ion Ion RNA-Seq LogRatio 2 1. Ion small RNA-Seq mirnaion PGM mirbasemirna TaqMan C t < 40 3 mirna2 mirna Ion PGM mirna Ion PGM mirna -3p -5p leading star * 2 hsa-mir-126 mirna Ion PGM 5' 3'isomir isomir 2. Ion PGM mirna -3p -5p TaqMan MicroRNA Ct log2mapped Ion PGM log2 hsa-mir-126 hsa-mir-126* TaqMan Not detected Ion PGM Total assays Intersection assays Intersection detected TaqMan GeneChip Array Ion Ion log 2 Ion PGM log 2 (/ ) PGM 316 PGM 314 Array 2. Ion small RNA-Seq Ion 316 Ion 314 ChipsLogRatio GeneChip mirna Ordering information Ion PGM PGM11-020S2 2 9,600,000 Ion PGM 1Torrent 1Ion OneTouch 2.0 Duo Ion PGM Ion PGM 1Torrent 1Ion Chef PGM11-C01S2 2 13,400,000 5

6 Ion Total RNA-Seq v2 RiboMinus Eukaryote v2 Ion Total RNA-Seq v2 small RNA small RNA small RNA 100ng RNA5ng mirna 1ng mirna small RNA 1 small RNA 16 RNA サンプル RiboMinus Eukaryote v2 RNA RNA RNA Total RNA18S 28S 5S 5.8S rrna 95rRNA 5 rrna/ PolyA+ mrna PolyA+ RNA RNA RNA RT-PCR rrna 95 1 RiboMinus Eukaryote System v2 1 5 g RNA Low Input RiboMinus Eukaryote System v2 100ng-1 g RNA 5 ライゲーション部位 5 アダプター 3 ライゲーション部位 3 アダプター アダプターのハイブリダイゼーション ライゲーション 15 分 30 分 目的の RNA サンプル ブロック 逆転写開始位置 逆転写 精製 30 分 1 時間 v2 1 RNA ライブラリの増幅 1 時間 精製 1 時間 合計 :4 時間 15 分 Ion Total RNA-Seq Step 1 Step 2 Step 3 Step 4 Hybridized RiboMinus probe Biotin Non rrnas Bead Streptavidin Biotin Non rrnas Magnet rrna-depleted RNA リボソーム RNAとプローブをハイブリダイゼーション 10 分 - 変性 20 分 - キャプチャー ストレプトアビジン - 磁気ビーズを用いて リボソーム RNA / プローブ複合体を捕捉 マグネットで複合体を吸着 クリーンナップ 30 分 Ion Total RNA-Seq v2 Ordering information Ion Total RNA-Seq kit v2 12Rxn ,000 Ion Total RNA-Seq kit v2 48Rxn ,000 Ion Xpress RNA-Seq Bc 01-16kit ,000 RiboMinus Eukaryote System v2 12 A ,700 Low Input RiboMinus Eukaryote System v2 12 A ,700 6

7 ERCC RNA Spike-In ERCC RNA Spike-In NIST National Institute of Standards & Technology ERCC External RNA Control Consortium RNA 92 RNA RNA polyasolid Total RNArRNA polyA RNA 10 6 PCR small RNA total RNA Spike-In rrna polya RNA RNA Spike-In RNA Spike-In 1 Spike-In 2 RNA PCR TaqMan Assay 1e+9 ERCC Mix 1(molecules/100ng mrna) 1e+8 1e+7 1e+6 1e+5 1e+4 1e+3 1e+2 1e+2 1e+3 1e+4 1e+5 1e+6 1e+7 1e+8 1e+9 ERCC Mix 2(molecules/100ng mrna) 3. ERCC ExFold RNA Spike-In ExFold RNA Spike-In polyA RNA ERCC RNA Spike-In 1 ExFold Spike-In Ordering information ERCC ExFold RNA Spike-In 10 µl ,000 ERCC RNA Spike-In 10 µl ,000 7

8 microrna TaqMan 1 microrna mirna mirna mirna mirna mirna TaqMan OpenArray MicroRNA Panel TaqMan Array Micro RNA CardMegaplex Primer Pools mirna 48 mirna 100 ng RNATaqMan PCR RT TaqMan OpenArray MicroRNA TaqMan Array MicroRNA1 100 ng RNA 4 ライブラリ作製 5 mirna mirbase mirna TaqMan Array MicroRNA Card 377miRNA 1 A B 377miRNA 2 TaqMan OpenArray MicroRNA Panel754miRNA 2 Megaplex RT Primers 377miRNA Applied Biosystems PCR Megaplex PreAmp Primer Pools mirna PCR TaqMan Universal PCR Master Mix II TaqMan OpenArray MicroRNA Panel AccuFill 3754TaqMan MicroRNA Assays TaqMan Array MicroRNA Card Card TaqMan MicroRNA Card A B 2 377miRNA Assay リアルタイム 1 cdna 増刷 cdnaの希釈サンプル調製 2 逆転写反応 (RT) PCR 装置による 6 ( オプション ) & ロードラン実行 データ解析 mir Vana TM mirna Isolation Kit TaqMan ABC Purification Kit mirna Megaplex TM RT Primers TaqMan MicroRNA Reverse Transcription Kit Megaplex TM PreAmp Primers & TaqMan PreAmp Master Mix TaqMan Array Card or OpenArray Plate QuantStudio 12K Flex, ViiA HT Fast リアルタイム PCR システム RealTime StatMiner Software DataAssist Software Total RNA xxxxxxxx xxxxxxxx xxxxxxxx xxxxxxxx xxxxxxxx xxxxxxxx xxxxxxxx xxxxxxxx xxxxxxxx xxxxxxxx xxxxxxxx xxxxxxxx xxxxxxxx xxxxxxxx 30 min 150 min 90 min 10 min 60 min 4. TaqMan OpenArray MicroRNA Panel TaqMan Array MicroRNA Cards mirnataqman OpenArray MicroRNA PanelsMegaplex TM PreAmp Primer Poolspre-amplification pre-amplification 4.25 TaqMan Array MicroRNA CardsMegaplex TM PreAmp Primer Poolspre-amplification

9 Megaplex Megaplex Primer Pools TaqMan OpenArray MicroRNA PanelTaqMan Array MicroRNA CardMegaplex Primer PoolsmiRBase mirna mirna Megaplex RT Primer Pools mirna Megaplex RT Primer Pools mirna RT cdnataqman MicroRNA Assays Megaplex PreAmp Primer Pools TaqMan Array MicroRNA Cards Megaplex PreAmp Primer Pools mirna Megaplex PreAmp Primer Pools 1 ng RNAmiRNA 5 mirna TaqMan MicroRNA TaqMan MicroRNA ArraysmicroRNA TaqMan MicroRNA Assay Megaplex Primer Pools TaqMan OpenArray MicroRNA Panels TaqMan OpenArray MicroRNA Panels 1 3 mirna Megaplex RT Primer PoolsMegaplex PreAmp Primer Pools TaqMan Universal PCR Master Mix II TaqMan Arrays MicroRNA Cards TaqMan OpenArray Panel log 4 log RNA PCR logmirna 1 mirna mirna TaqMan 4 RT mirna RNA mirna 1 g 100 ng in silico mirna 1 1,000 ng in silico Megaplex Primer Pools TaqMan Array Cards Megaplex Primer Pools Megaplex Human A Pool の検出効率 Megaplex Human B Pool の検出効率 検出 % 検出 % ntc 0.1 fm 1 fm 10 fm 100 fm 1 pm 10 pm 100 pm 0 ntc 0.1 fm 1 fm 10 fm 100 fm 1 pm 10 pm 100 pm RT PreAmp テンプレート濃度 RT PreAmp テンプレート濃度 5. Megaplex Primer Pools TaqMan Array MicroRNA Cards mirna TaqMan MicroRNA Array A Card B Card Card mirna 10 pmtotal RNA 10ng/ L 6 log 4 log Megaplex NTC 9

10 Ordering information microrna mirvana mirna Isolation Kit TaqMan mirna ABC Purification Kit microrna RT TaqMan MicroRNA RT Kit TaqMan MicroRNA RT KitTaqMan MicroRNA AssaysMegaplex Primer PoolsTaqMan MicroRNA Assays RT TaqMan MicroRNA RT Kit ,300 TaqMan MicroRNA RT Kit ,800 Megaplex Primer Pools Megaplex RT Primers stem-loopedtaqman MicroRNA Arrays mirna mirna RNA Megaplex RT Primers Pool for Human Megaplex RT Primers, Human Pool A ,900 Megaplex RT Primers, Human Pool B v ,900 Megaplex RT Primers, Human Pool Set v3.0 RT Primers Pools A&B ( & ) ,000 Megaplex RT Primers Pool for Rodent Megaplex RT Primers, Rodent Pool A ,900 Megaplex RT Primers, Rodent Pool B v ,900 Megaplex RT Primers, Rodent Pool Set v3.0 RT Primers Pools A&B( & ) ,000 Megaplex Primer Pools Pool APool BTaqMan MicroRNA Arrays A B PreAmpcDNA Megaplex PreAmp Primers1ngRNA Megaplex PreAmp Primers Pool for Human Megaplex PreAmp Primers, Human Pool A ,900 PreAmp Megaplex PreAmp Primers, Human Pool B v3.0 PreAmp Megaplex PreAmp Primers, Human Pool Set v3.0 PreAmp Primers Pools A&B ( & ) , ,000 Megaplex RT and PreAmp Primers Package for Human Megaplex Primer Pools, Human Pools A ,200 RT Primers( ) + PreAmp Primers( ) Megaplex Primer Pools, Human Pools B v ,200 RT Primers( ) + PreAmp Primers( ) Megaplex Primer Pools, Human Pools Set v3.0 RT Primers Pools A&B ( (= & )) PreAmp Primers Pools A&B ( (= & )) ,400 10

11 Megaplex PreAmp Primers Pool for Rodent Megaplex PreAmp Primers, Rodent Pool A ,900 PreAmp Megaplex PreAmp Primers, Rodent Pool B v ,900 PreAmp Megaplex PreAmp Primers, Rodent Pool Set v3.0 PreAmp Primers Pools A&B( & ) ,000 Megaplex RT and PreAmp Primers Package for Rodent Megaplex Primer Pools, Rodent Pools A ,200 RT Primers ( ) + PreAmp Primers ( ) Megaplex Primer Pools, Rodent Pools B v ,200 RT Primers( ) + PreAmp Primers( ) Megaplex Primer Pools, Rodent Pools Set v3.0 RT Primers Pools A&B( (= & )) PreAmp Primers Pools A&B( (= & )) ,400 TaqMan PreAmp Master Mix TaqMan PreAmp Master Mix ,000 11

12 mirnapcr Human TaqMan Array Human MicroRNA Cards TaqMan Array Human MicroRNA A Card v ,000 TaqMan Array Human MicroRNA A+B Cards Set v ,600 A array ( ) + B Array ( ) 4 TaqMan Array Human MicroRNA B Card v ,000 Rodent TaqMan Array Rodent MicroRNA Cards TaqMan Array Rodent MicroRNA A Card v ,000 TaqMan Array Rodent MicroRNA A+B Cards Set v ,600 A array ( ) + B Array ( ) 4 TaqMan Array Rodent MicroRNA B Card v ,000 TaqMan MicroRNA Panels /Plate /Plate TaqMan OpenArray Human MicroRNA Panel QuantStudio TM 12K Flex * 100,000 TaqMan OpenArray Rodent MicroRNA Panel QuantStudio TM 12K Flex * 100,000 TaqMan OpenArray Real-Time Master Mix, 1.5mL 1.5 ml ,200 TaqMan OpenArray Real-Time Master Mix, 5mL 5 ml ,500 QuantStudio 12K Flex OpenArray Accessories Kit ,900 * QuantStudio 12K Flex PCR PCR QuantStudio 12K Flex PCR Applied Biosystems Viia TM 7 PCR 12

13 mirna mirna mirna mirna TaqMan PCR mirna TaqMan MicroRNA Assays mirna ~22 nt mirna mirnamirna 3 RT/miRNA RT TaqMan PCR C. eleganssanger TaqMan MicroRNA Assays mirbase C. elegansmirbase mirna mirna TaqMan Assays mirbase mirna TaqMan MicroRNA Assays mirna 3'cDNA TaqMan PCR mirna TaqMan (RT) MicroRNA PCR 4 5 Assay microrna microrna Stem-loop primer 30 min 65 min 25 min 90 min mirvana mirna Isolation Kit TaqMan mirna ABC Purification Kit RecoverAll Total Nucleic Acid Isolation Kit for FFPE TaqMan MicroRNA Cells-to-CT Kit TaqMan MicroRNA Reverse Transcription Kit TaqMan MicroRNA Assays TaqMan Universal Master Mix II TaqMan MicroRNA Assays Applied Biosystems PCR Instrument software (included) RQ Manager enterprise software (optional) RealTime StatMiner Software (optional) 13

14 TaqMan MicroRNA Assays 2 TaqMan MicroRNA Assays TaqMan Sanger mirbase mirna Sanger mirbase TaqMan MicroRNA Assays Custom TaqMan Small RNA Assays mirnataqman mirna Assay 200 Custom TaqMan Small RNA Assay RNA mirna pirna piwiinteracting RNA snrna snorna small RNA C. elegans RNA Endogenous controls for real-time quantitation of mirna using TaqMan MicroRNA Assays TaqMan mirna Target Sequence Stem-loop RT primer mirna ng RNA 7 log 1 mirna 7 Forward Primer Reverse Transcription Real-Time PCR 2 PCR Q TaqMan probe F Reverse primer 6. TaqMan MicroRNA Assay 2 mirna PCR 14

15 Mature mirnas Precursors mirna Assays Perfectly Mismatched C t C t (mismatch matched C t vs. match) C t C t (Precursor vs. mature) let-7a Looped Linear mirna mirna Stem-loop RT primer Linear RT primer 7. TaqMan MicroRNA Assays mirna let-7a RT TaqMan MicroRNA AssayRT/PCR/TaqMan let-7a let-7e let-7a 2miRNA 1.5 x 10 8 mirna let-7a TaqMan MicroRNA Assay RTRTPCR RT mirna mirna mirna Rn 0.1 C T 25 70M copies 7 copies Slope = 3.36 R 2 = , Cycle Panel A. Amplification Plot Quantity Panel B. Standard Curve 8. TaqMan MicroRNA Assays 7 log 1 mirna mirna TaqMan MicroRNA Assays lin-4 mirna lin-4 TaqMan MicroRNA Alin-4 mirna7 PCR RNA1.3 x 10-3 fm71.3 x 10 4 fm7 x 10 7 B lin-4 mirna 15

16 Ordering information microrna mirvana mirna Isolation Kit TaqMan mirna ABC Purification Kit microrna RT TaqMan MicroRNA RT Kit TaqMan MicroRNA RT KitTaqManR MicroRNA AssaysTaqMan MicroRNA Assays RT TaqMan MicroRNA RT Kit ,300 TaqMan MicroRNA RT Kit 1, ,800 mirnapcr TaqMan MicroRNA Assays TaqMan MicroRNA AssaysPCR microrna mirnamirna TaqMan MicroRNA Assays, Inventoried ,800 mirnart primer 5X reaction mix(50 / 15 µl) mirnataqman Assay 20X reaction mix (150 / 20 µl) Arabidopsis thaliana Caenorhabditis elegans Drosophila melanogaster TaqMan MicroRNA Assays, Made-to-Order, Extra Small mirnart primer 5X reaction mix(25 / 15 µl) mirnataqman Assay 20X reaction mix (75 / 20 µl) ,600 TaqMan MicroRNA Assays, Made-to-Order, Small mirnart primer 5X reaction mix(50 / 15 µl) mirnataqman Assay 20X reaction mix (150 / 20 µl) ,000 TaqMan MicroRNA Assays, Made-to-Order, Medium ,800 TaqMan MicroRNA Assays, Made-to-Order, Large 2,900 2, ,500 Custom TaqMan Small RNA Assays * Custom TaqMan Small RNA Assays, X ,800 Custom TaqMan Small RNA Assays, ,000 Custom TaqMan Small RNA Assays, 750 2, ,200 Custom TaqMan Small RNA Assays, 2,900 8, ,600 *20µL 16

17 TaqMan Universal Master Mix II TaqMan Universal Master Mix IITaqManTaqMan MicroRNA Assays PCR TaqMan Universal Master Mix II, with UNG イ * TaqMan Universal Master Mix II, with UNG 1mL ,000 TaqMan Universal Master Mix II, with UNG 5mL ,000 TaqMan Universal Master Mix II, with UNG 2 x 5mL ,000 TaqMan Universal Master Mix II, with UNG 5 x 5mL 1, ,000 TaqMan Universal Master Mix II, with UNG 10 x 5mL 2, ,000 TaqMan Universal Master Mix II, with UNG 50mL 2, ,000 TaqMan Universal Master Mix II, no UNG イ * TaqMan Universal Master Mix II, no UNG 1mL ,000 TaqMan Universal Master Mix II, no UNG 5mL ,000 TaqMan Universal Master Mix II, no UNG 2 x 5mL ,000 TaqMan Universal Master Mix II, no UNG 5 x 5mL 1, ,000 TaqMan Universal Master Mix II, no UNG 10 x 5mL 2, ,000 TaqMan Universal Master Mix II, no UNG 50mL 2, ,000 *50µL デジタル PCR システム リアルタイム PCR システム QuantStudio 3D デジタル PCR システム QuantStudio 12K Flex リアルタイム PCR システム QuantStudio 7K Flex リアルタイム PCR システム QuantStudio 6 Flex リアルタイム PCR システム 17

18 microrna mirna mirna mirna mimic gain-of-function loss-of-function mirna inhibitor down-regulation mirnaup-regulationdown-regulation mirna Ambion 1 mirnaambion microrna mimic inhibitor mirna Ambion mirvana mirna mimic mirvana mirna inhibitor mirna gain-of-function loss-offunction sirna gain-of-function loss-of-functionmirna mirna PCR mirvana mirna mimicmirvana mirna inhibitor mirna mrna mirna mirna / mrna mirna mirna mimic inhibitor mrna 3' 5' mirvana mirna mimic 3' 5' mirvana mirna Inhibitor mirvana mirna mimics mirvana mirna inhibitors Lipofectamine RNAiMAX Transfection Agent Neon Transfection System TaqMan Gene Expression Cells-to-CT Kit TaqMan Gene Expression Assays Applied Biosystems PCR NuPAGE iblot MagicMark XP mirna In vitro In vivo mirna mirvana mirna mimics & inhibitors Lipofectamine RNAiMAX mirna mirvana mirna mimics & inhibitors, in vivo ready Invivofectamine

19 microrna mirvana mirna mimics & inhibitors: mirbase in vitro in vitroin vivo mrna upregulation % mir 122 mirna122 = mir122 mirna inhibitor NT =Not transfected Neg1 = Negative Control #1 NT AldoA NEG1 mir 122 NT NEG1 mir NT NEG1 mir NT Hfe2 Slc35a4 Lass6 NEG1 mirvana mirna mimics mirvana mirna mimics RNA sirna mirna mirna RISC RNAinduced silencing complex8 sirna mirna Dose response mirvana mirna Inhibitors mirvana mirna Inhibitor mirna mirna inhibitor mirvana mirna Inhibitor 4 mirna in vivo in vivo Ready mirnain vivo TaqMan mirvana mrna inhibitor TaqMan 10 mimic inhibitor in vivo 1 mimic inhibitor 10. mirvana mirna inhibitor in vivomirna mir122negative Control #1 mirvana mirna inhibitor Invivofectamine 2.0Balb-C 37 mg/kg24 mir122 4 TaqMan MicroRNA Assay Negative ControlmiR122 4 mrna Fold change mirna mimic Neg reporter 1 mirna mimic Neg mirna mimic 11. mirvana mirna mimic mirna mature star mature star 6miRNA mirvana mirna mimic mature Negative Control 510 star Negative Control mirna Neg mirna mimic mirna mirna mirvana mirna mimicinhibitor nmol 50 nm 50mirVana mirna mimicinhibitor Positive Control Negative Control Neg mirna mimic Neg mirna mimic reporter 2 reporter 3 reporter 4 reporter 5 reporter 6 FW RV Neg mirvana mirna mimics 3' 5' 3' 5' 5'Cap AAAAAAAAAAAAA3' 3' 5' No translation mirna mimic Sequence-specific loading into RISC-like complex mirvana mirna inhibitors mrna 3' 5' 3' 5' 5'Cap AAAAAAAAAAAAA3' Translation mirna inhibitor Complementary binding of specific mirnas no RISC-like complex association 3' 5' 9. mirvana mirna mimics mirvana mirna inhibitors 19

20 Ordering information microrna microrna mirvana mirna mimic mirvana mirna mimic 5 nmol ,000 mirvana mirna mimic 20 nmol ,000 mirvana mirna mimic, in vivo Ready 250 nmol ,000 Customer-defined mirvana mirna Mimic 5 nmol ,000 Customer-defined mirvana mirna Mimic 20 nmol ,000 Customer-defined mirvana mirna Mimic, in vivo Ready 250 nmol ,000 mirvana mirna Mimic Negative Control #1 5 nmol ,000 mirvana mirna Mimic Negative Control #1 20 nmol ,000 mirvana mirna Mimic Negative Control #1, in vivo Ready 50 nmol ,000 mirvana mirna Mimic Negative Control #1, in vivo Ready 250 nmol ,000 mirvana mirna Mimic mir-1 Positive Control 5 nmol ,000 mirvana mirna Mimic mir-1 Positive Control 20 nmol ,000 mirvana mirna Mimic mir-1 Positive Control, in vivo Ready 50 nmol ,000 mirvana mirna Mimic mir-1 Positive Control, in vivo Ready 250 nmol ,000 mirvana mirna mimic mirbase v 20* 0.25 nmol each mirvana mirna mimic mirbase v 20* 1 nmol each * mirvana mirna inhibitor mirvana mirna inhibitor 5 nmol ,000 mirvana mirna inhibitor 20 nmol ,000 mirvana mirna inhibitor, in vivo ready 250 nmol ,000 Customer-defined mirvana mirna Inhibitor 5 nmol ,000 Customer-defined mirvana mirna Inhibitor 20 nmol ,000 Customer-defined mirvana mirna Inhibitor, in vivo ready 250 nmol ,000 mirvana mirna Inhibitor Negative Control #1 5 nmol ,000 mirvana mirna Inhibitor Negative Control #1 20 nmol ,000 mirvana mirna Inhibitor Negative Control #1, in vivo Ready 50 nmol ,000 mirvana mirna Inhibitor Negative Control #1, in vivo Ready 250 nmol ,000 mirvana mirna Inhibitor let-7 Positive Control 5 nmol ,000 mirvana mirna Inhibitor let-7 Positive Control 20 nmol ,000 mirvana mirna inhibitor mirbase v 20* 0.25 nmol each mirvana mirna inhibitor mirbase v 20* 1 nmol each *

21 mirna& Lipofectamine RNAiMAX reagent sirna RNAimiRNA mimic mirna inhibitor Neon Transfection System Neon sirna/mirna sirna Invivofectamine 2.0 sirna mirvana mirna mimics and inhibitors, in vivo readyin vivo mirna mirna& Lipofectamine RNAiMAX 12. Neon Transfection Neon Lipofectamine RNAiMAX 0.1 ml ,900 Lipofectamine RNAiMAX 0.3 ml ,000 Lipofectamine RNAiMAX 0.75 ml ,000 Lipofectamine RNAiMAX 1.5 ml ,500 Lipofectamine RNAiMAX 15 ml ,000 Invivofectamine 2.0 In vivo Invivofectamine 2.0 Reagent Starter Kit 1mL ,000 Invivofectamine 2.0 Reagent 5mL ,000 Starter kitsirna Ambion In Vivo FVll sirna50nmol Neon Transfection System Neon Transfection System* 1 1 MPK ,000 Neon Transfection System Starter Pack* 2 1 MPK5000S 1,050,000 1 Neon Transfection System(MPK5000) Neon Neon Pipette MPP100Neon Pipette Station MPS100 2 Starter Pack MPK5000S Neon Transfection System(MPK5000) Transfection Kit(MPK10096 MPK10961 Neon Transfection System Neon Transfection System Kit (100 L) 1 Neon 96 MPK ,000 1 Neon 25 MPK ,000 Neon Transfection System Kit (10 L) 1 Neon 96 MPK ,000 1 Neon 25 MPK ,000 Neon Transfection Tubes 1 (100 ) MPT100 49,000 Neon Pipette MPP100 75,000 Neon Pipette Station MPS ,000 21

22 mirna mirnagain-of-function loss-of-function mrna micrornamrna PCR TaqMan PCR SYBR PCR RNA microrna 22

23 microrna mirna small RNA small RNA <200 nt mirna New! TaqMan mirna ABC Purification Kit TaqMan mirna ABC anti-mirna Bead Capture Purification Kit TaqMan mirna Assay TaqMan Array Cardanti-miRNA FFPE mirna 10 L 50 LmiRNA PCR TRIzol 75 RNA TaqMan mirna ABC Purification KitmiRNA 21Human Panel A Human Panel B TaqMan Array Human MicroRNA A Card B Card 754 mirnamirna mirna mirvana mirna Isolation Kit Lysis Solution RNaseRNA RNA RNA RNA mirna mirnasmall RNA small RNA RNA small RNA <200 nt RecoverAll Total Nucleic Acid Isolation Kit FFPE RecoverAll Kit 451.5FFPE RNA FFPE RNA DNA RNA PCR TaqMan MicroRNA Cells-to-CT Kit 10 PCR RNA PBS 8 DNase Stop Solution TaqMan MicroRNA Cells-to-CT Kit 10 TaqMan MicroRNA TaqMan Universal PCR Master Mix II TaqMan mirna 23

24 microrna mirvana mirna Isolation Kit mirvana PARIS Kit MagMAX -96 for Microarrays Total RNA Isolation Kit TaqMan MicroRNA Cells-to-C T Kit TaqMan mirna ABC Purification Kit RecoverAll Total Nucleic Acid Isolation Kit for FFPE Tissues TRI Reagent Cells-to-C T DNase RNA PCR Anti-miRNA mirna mg mg 5x mg L mg FFPE5 10 m 50 L FFPE 20 m RNase small RNA mirna 30 RNA small RNA small- RNA mrna mirna sirna small RNA 10 PCR TaqMan Micro RNA Array Card Panel TRIzol 75 RTqPCR TaqMan Human MicroRNA FFPE DNA mirna Proteinase K PCR >50% PCR PCR 24

25 Ordering information microrna micrornatotal RNA mirvana mirna Isolation Kit mirvana mirna Isolation Kit 40 AM ,300 mirvana mirna Isolation Kit without Phenol 40 AM ,000 mirna mirvana PARIS Kit mirvana PARIS Kit 40 AM ,800 MagMax MagMAX Total Nucleic Acid Isolation Kit 100 AM ,800 MagMAX -96 Blood RNA Isolation Kit 96 AM ,800 MagMAX -96 Total RNA Isolation Kit 96 AM ,500 MagMAX Viral RNA Isolation Kit 50 AM ,700 MagMAX -96 Viral RNA Isolation Kit 96 AM ,800 RNA Single Place Magnetic Stand 1 AM ,400 Magnetic Stand-96 1 AM , well Magnetic-Ring Stand 1 AM ,000 TaqMan MicroRNA Cells-to-C T Kit RNA microrna mirna * TaqMan MicroRNA Cells-to-C T Kit ,000 * microrna TaqMan mirna ABC Anti-miRNA Bead Capture Purification Ki TaqMan mirna ABC Purification Kit-Human Panel A ,000 TaqMan mirna ABC Purification Kit-Human Panel B ,000 25

26 Total Exosome Isolation RNA エキソソーム回収 Total Exosome Isolation 各サンプルに対応した 5 種類の専用試薬 超遠心を使わずに短時間でエキソソームを回収 対応サンプルタイプ 細胞培養上清血清血漿尿脳脊髄液腹水羊水母乳唾液 RNA とタンパク質の精製 解析 特定の抗原を持つエキソソームの単離 フロー解析 エキソソームの蛍光標識 Total Exosome RNA & Protein Isolation Kit 同一サンプルからタンパク質と RNA を回収可能 Exosome Immunoprecipitation 磁気ビーズを利用した わずか 30 分のシンプルなプロトコールウェスタン解析用抗体エキソソームマーカータンパク質のウェスタン解析用抗体 Exosome-Human Isolation/Detection (CD9, CD63, CD81, Ep-CAM) 高純度の CD9, CD63, CD81, Ep-CAM, 陽性エキソソームを単離 Exosome-Streptavidin for Isolation/Detection 前濃縮したエキソソームから作業時間 1 時間以内に特定の抗原を持つエキソソームを単離可能 SYTO RNASelect Green Fluorescent Cell Stain BODIPY TR ceramide エキソソーム中の RNA もしくはエキソソームの膜を蛍光標識 Exosome Spin Columns (MW 3000) 蛍光標識したエキソソームの精製 Total Exosome Isolation Total Exosome Isolation 50 ml ,000 Total Exosome Isolation 6 ml ,000 Total Exosome Isolation 6 ml ,000 Total Exosome Isolation 50 ml ,000 Total Exosome Isolation 6 ml ,000 Exosome-Human CD63 Isolation/Detection 3 L 30 prep/150 rxn 10606D 40,000 Exosome-Human CD9 Isolation 2mL 50 prep 10614D 58,000 Exosome-Human CD81 Isolation 2mL 50 prep 10616D 58,000 Exosome-Human Ep-CAM Isolation 2mL 50 prep 10618D 58,000 Exosome-Human CD9 Flow Detection 2mL 100 rxn 10620D 26,000 Exosome-Human CD81 Flow Detection 2mL 100 rxn 10622D 26,000 Exosome-Human Ep-CAM Flow Detection 2mL 100 rxn 10624D 26,000 Exosome-Streptavidin for Isolation/Detection* 3mL 30 prep/150 rxn 10608D 40,000 RNA Total Exosome RNA & Protein Isolation Kit 40 rxn ,000 Exosome Immunoprecipitation Protein A * 1 ml 10610D 22,000 Exosome Immunoprecipitation Protein G * 1 ml 10612D 22,000 Exosome Spin Columns MW ,000 SYTO RNASelect Green Fluorescent Cell Stain 100 µl S ,900 BODIPY TR Ceramide 250 µg D ,900 Exosome Anti-CD9 0.2 ml 10626D 29,000 Exosome Anti-CD ml 10628D 29,000 Exosome Anti-CD ml 10630D 29,000 * 26

27 TaqMan Assays ID TaqMan MicroRNA Assays mirvana mirna mimics mirvana mirna inhibitors ご登録からご注文までの流れ ウェブ登録画面に必要事項を記入する Applied Biosystems 製品取扱代理店に登録用紙を渡す 代理店が必要事項を記入し弊社宛に郵送 ユーザー ID とパスワードをご登録者に でお知らせ ログインしてオンラインオーダー可能

28 References 1. Bartel DP (2004) MicroRNAs: Genomics, biogenesis, mechanism, and function. Cell 116(2): Soifer HS, Rossi JJ, Saetrom P (2007) MicroRNAs in disease and potential therapeutic applications. Mol Ther 15(12): Griffi ths-jones S, Saini HK, van Dongen S, Enright AJ (2008) mirbase: tools for microrna genomics. Nucleic Acids Res 36: D154 D158. facebook.com/lifetechnologiesjapan@lifetechjpn For Research Use Only. Not for use in diagnostic procedures Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientific and its subsidiaries unless otherwise specified. Printed in Japan. GNL130-B1407OB TEL FAX TEL FAX