NEXT No.36

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1 2016 / July No. 36 Science, Products, and Information NEXT Interview TiPS P. 04 Arcturus XT Laser Capture Microdissection System P. 05 Ion AmpliSeq Panel P. 06 NanoDrop & Qubit 3.0 Fluorometer P. 08 ADS P. 09 QuantStudio 3D PCR P. 10 Essencial 8 Flex / TaqMan hpsc Scorecard Panel P. 12 P. 13 Lipofectamine CRISPRMAX Cas9 Transfection Reagent P. 15 Click-iT Colorimetric IHC Detection Kit P. 16 EVOS FL Auto 2 Imaging System P. 18 N-PER Neuronal Protein Extraction Reagent Syn-PER Synaptic Protein Extraction Reagent

2 Interview T ips T T 33 T TiPS T ips TiPS ips T ES ips ES ips ips 1 2 T 40 ips 2013 TiPS T HLA HLA T ADA 1994 T page 02 page 03

3 Ion AmpliSeq Panel User's Voice NGS DNA Ion AmpliSeq % BRCA1/2 BRCA1/ BRCA AmpliSeq Ion AmpliSeq Ion AmpliSeq Designer PCR BRCA1/ % 610 Ion Proton BRCA1/2 BRCA1/ ATM MRE11A MSH63 3 BRCADNA MSH6 DNA BRCA1/2 DNA PARP FDA BRCA1/2 FFPE Arcturus XT Laser Capture Microdissection System Heterogeneity Applied Biosystems Arcturus XT LCM System LCM Ion AmpliSeq Designer DNA LCM 3 DNA Streamline Multigene panel analysis identified germline mutations of DNA repair genes in breast and ovarian cancer Yosuke Hirotsu et al. Molecular Genetics & Genomic Medicine (2015); 3(5): Ion AmpliSeq Panel PCR Ion Torrent 6,144 / 1 5 Mb DNA10ng / 1 FFPE Ion AmpliSeq DNA Cancer Hotspot Panel v2 50 Comprehensive Cancer Panel 409 Ion AmpliSeq RNA Colon & Lung Cancer Panel v2 BRCA1 & BRCA2 Panel TP53 Panel AML Panel Inherited Disease Panel 328/700 Hearing Loss Research Panel v1 Dementia Research Gene Panel20 Precision ID Identity / Ancestry Panel DNA / 2 Pharmacogenomics Panel 40 TB Research Panel8 RNA Cancer Panel Cancer Hotspot Panel v2 RNA Apoptosis Panel 267 RNA Fusion Lung Cancer Research Panel 70 Ion Torrent RNA Custom Panel 20,000 RNA Gene Fusion designs 1,000 Ebola Research Panel Cancer Hotspot Panel v2 2,790 COSMIC Mutations KRAS, BRAF, EGFR Comprehensive Cancer Panel 4,465 COSMIC Mutations AR, NFKB2, SOX ,992 RNA Cancer Panel KRAS, BRAF, EGFR RNA Apoptosis Panel c-myc, TP53, FAS, BAX TP53 Panel TP BRCA1 and BRCA2 Panel BRCA1, BRCA Colon & Lung Cancer Research Panel v2 NRAS, PTEN, PIK3CA AML Research Panel DNMT3A, RUNX RNA Fusion Lung Cancer Research Panel ALK, RET, ROS1,NTRK Arcturus XT LCM SystemIR UV ArcturusXT IR & UV 1 1 ARCTURUSXT-B-S2 24,900,000 ArcturusXT IR & UV 1 1 ARCTURUSXT-BF-S2 28,400,000 Ion AmpliSeq Cancer Hotspot Panel v ,000 Ion AmpliSeq Comprehensive Cancer Panel ,000 Ion AmpliSeq Inherited Disease Panel ,000 Ion AmpliSeq Custom Panel 7503, page 04 page 05

4 Technical Review NanoDrop & Qubit 3.0 Fluorometer 2 User's Voice 2 UV Thermo Scientific NanoDrop Invitrogen Qubit Fluorometer NanoDrop µl Qubit 3.0 Fluorometer SNP 1 NanoDropQubit NanoDrop One Acclaro Qubit 3.0 Fluorometer FFPE DNA Qubit 0.4ng / µl 1 Qubit NanoDrop One NanoDrop One 1 A ,750,000 NanoDrop One C 1 A ,050,000 Qubit 3.0 Fluorometer 1 Q ,000 Qubit 3.0 Quantitation Starter Kit 1 Q ,100 Qubit 3.0 NGS Starter Kit 1 Q ,100 Platinum SuperFi DNA Polymerase PCR 3 Invitrogen Platinum SuperFi DNA Polymerase Taq DNA Polymerase 100 DNA polymerase High- Fidelity Long PCR(>20Kb) GC-rich >75%PCR PCR Platinum SuperFi DNA Polymerase PCR 10Kb Long PCR 10Kb PCR 1 Tm PCR Tm Tm Tm Calculator com/tmcalculator primer Tm 1 1 Tm Calculator Primer PCR primer20nt primer set Tm primer Tm3 10 primer20nt primer set Tm 10 Point 1 2-step protocol primer Tm 69 20nt72 20nt Tm 72 primer set 72 / 1 Platinum SuperFi DNA Polymerase PCR Point 2 Point 3 PCR Platinum SuperFi DNA Polymerase ,000 Platinum SuperFi Green DNA Polymerase ,000 Platinum SuperFi PCR Master Mix ,500 Platinum SuperFi Green PCR Master Mix ,500 dntp dutp ditpdntp primer DNA DNA PCR Primer Primer SuperFi GC GC-rich targets >65% GC PCR 2-step protocol Primer Primer Primer Primer page 06 page 07

5 ADS! User's Voice! ADS,TaqMan Gex A ,000 StepOne, StepOnePlus, 7000, 7300,7500, ADS,Custom TaqMan GEx A , Fast, 7900HT, ViiA7, ADS,Multiplex 2Dye TaqMan Gex A ,000 QuantStudio 3 / 5 / 6 / 7 / 12K ADS Sequencing A , / 3130xl, 3500 / 3500xL,3730 / 3730xl NEW! ADS,TaqMan GEx with Training A ,000 StepOne, StepOnePlus, 7000, 7300,7500, ADS,Custom TaqMan GEx with Training A , Fast, 7900HT, ViiA7, ADS,Multiplex 2Dye TaqMan GEx with Training A ,000 QuantStudio 3 / 5 / 6 / 7 / 12K ADS,Multiplex 3Dye TaqMan GEx with Training A ,800,000 AADS,Multiplex 4Dye TaqMan GEx with Training A ,800,000 ADS Sequencing with Training A , / 3130xl, 3500 / 3500xL,3730 / 3730xl BRAF PCR 7-9BRAF 1 QuantStudio 3D PCR BRAF 1 HE PCR 0.17% QuantStudio 3D PCR 1 RFLP PCR PCR 2 PCR 50 BRAF100% 2 PCR TaqMan PCR PCR TanqMan PCR 2 BRAF 4 PCR 13,2% RFLP 0.52% PCR6.8% PCR 0.14% 2016 JCVIM digital PCR BRAF Liquid biopsy 9 DNA QOL PCR20,000 QuantStudio 3D PCR ProFlex PCR 1 QS3D-PF 2 6,400,000 Custom TaqMan SNP Genotyping Assays for Rare Mutation Analysis , ,900 page 08 page 09

6 Essential 8 TaqMan hpsc ScoreCard Panel Design Space DS PATQbD DS ips ESiPS CMOS TaqMan hpsc Scorecard Panel QuantStudio 12K Flex PCR Quality by Testing LC/MS QbT TaqMan hpsc PAT ScoreCard Panel Gibco Essential 8 PCR design AppliedBiosystems TaqMan Quality by Design QbD hpsc ScoreCard Panel Essential 8 8 QbD BSA CMOS PAT Process Analytical Testing PAT PlosOne PAT N PAT QbD Gibco VTN-N 93 Essential 8 Flex ips/es TaqMan hpsc Scorecard Panel ES/iPS Gibco Essential 8 Flex ips/es 8 BSA Applied Biosystems TaqMan hpsc Scorecard Panel ES/iPS93 Applied Biosystems PCR Alex Meissner 2011 Cell Essential 8 Flex Medium 500 ml A ,000 Vitronectin Truncated Recombinant Human VTN-N 1 ml A ,000 TaqMan hpsc Scorecard Kit, Fast 96-well plate 2 1 A ,000 TaqMan hpsc Scorecard Panel, Fast 96-well 2 1 A ,500 page 10 page 11

7 ! User's Voice NEW! CRISPR-Cas9 ips 2 T7 Strings ORF vector ips ips mrna Vector ips ips RNPLipofectamine CRISPRMAX HCT116 HepG2 Cas9 grna in vitro DNAgRNA Cas9 grna Invitrogen GeneArt Genomic Cleavage Detection Kit Cas9 DNA CRISPR-Cas9 PCRin vitro ZFN TALEN CRISPR-Cas9 Cas9 Cas9 grna RNA Cas9 mrna Cas9 grna RNP Ribonucleo-protein DNA 500~1000 RNP DNA Lipofectamine CRISPRMAX PITCh Precise Integration into Target Chromosome Cas9 DNA 320 Cas9 DNA Lipofectamine CRISPRMAX Cas9 Transfection Reagent! T7 Strings 1 A31794JP 462,000 ORF vector 1 A31793JP 383,000 ips 1 A31795JP 511,000 ips 1 A31796JP 321,000 1 A31791JP 235,000 1 A31792JP 197,000 Invitrogen Lipofectamine CRISPRMAX Cas9 Transfection Reagent Cas9 Nuclease RNA Lipofectamine Lipofectamine CRISPRMAX Cas9 Transfection Reagent 0.1mL CMAX ,800 GeneArt Platinum Cas9 nuclease Cas9 10 µg 1µg/µL B , µg 1µg/µL B ,000 GeneArt Genomic Cleavage Detection Kit 20 A ,100 page 12 page 13

8 Technical Review CellInsight CX7 HCS ATG CellInsjght CX7 HCS CX7A1110 Click-iT Colorimetric IHC Detection Kit Invitrogen Click-iT EdU Colorimetric IHC Detection Kit Invitrogen Click-iT TUNEL Colorimetric IHC Detection Kit HAP-1CRISPR ATG5 ATG5 KO WT Thermo Scientific CellInsight CX7 HCS 24, KO NEW! HAP-1ATG5Invitrogen GeneArt CRISPR LC-3 Invitrogen NuclearMask Blue Invitrogen ClickiT EdU Invitrogen CellROX Green Invitrogen Click-iT OPP Invitrogen LysoTracker Red Invitrogen CellEvent Green 1 LC-3 CellInsight CX7 LC KOWT Click-iT EdU Colorimetric IHC Detection Kit 5-ethynyl-2'-deoxyuridine EdU DNA 1 LC LC-3 WT KO Click-iT EdU Alexa Fluor 488 kit Click-iT EdU Colorimetric IHC Detection Kit 2 EdU BrdU Click-iT EdU Colorimetric kit BrdU Staining kit EdU 3BrdU 23 LysoTracker RedCellInsjghtCX7 KOWT KOWT 3 KOWTLysoTracker RedCellInsjght CX7 Click-iT TUNEL Colorimetric IHC Detection Kit TUNEL DNA dutp ATG 11, ,000 CellInsjght CX Invitrogen ClickiT Plus TUNEL Assay for In Situ Apoptosis Detection, Alexa Fluor 647 DAPI Click-iT TUNEL Colorimetric IHC Detection Kit Click-iT EdU Colorimetric IHC Detection Kit 50 C ,800 Click-iT TUNEL Colorimetric IHC Detection Kit 1 50 C ,000 page 14 page 15

9 EVOS FL Auto 2 Imaging System NEW! Technical Review Attune NxT Flow Cytometer Z- Invitrogen EVOS FL Auto 2 Imaging System Invitrogen Attune NxT Flow Cytometer (10 20 µl/min) 100 1,000 µl/min 2MHz XY 4 Thermo Scientific Micro Studio Image Analysis Software Invitrogen EVOS Onstage Incubator CD3 CD4 CD8 CD19 CD45 CD56 6 1T TBNK Attune NxT µlcd2 ml High-Yield Lyse Fixative-Free Lysing Solution ( HYL250) (A) CD45 CD45+ (B) CD3 CD3+ TCD3 B NK C CD3+ CD4 CD8 CD4+ T CD4+, CD3+, CD45+ CD8+ T CD8+, CD3+, CD45+(D) CD3 CD56 CD19 CD56 NKCD19 B /µl Invitrogen LIVE/DEAD BacLight 1 40xBPAE DAPI Invitrogen Texas Red -X Phalloidin anti ß-tubulin BODIPY FL goat anti mouse IgG 2 40x Invitrogen Click-iT EdU Colorimetric IHC Detection Kitp15 Bacterial Viability Kit Attune NxT 3 3 SYTO 9propidium iodide (PI) (LIVE/DEAD BacLight Bacterial Viability KitL7012) 1: L/min 488 nm SYTO 9 530/30 nm BL1 PI 695/40 nm BL3 SYTO 9 PISYTO 9PI 1 /µl 3 /µl EVOS FL Auto 2 4 Ph, 10 Fl, 20 Fl, 40 Fl XY GFP, RFP, DAPI & 7,500,000 Attune NxT Flow Cytometer Blue/Red/Violet/Yellow Lasers 1 A ,950,000 EVOS Onstage Incubator 1 AMC1000 1,750,000 LIVE/DEAD BacLight Bacterial Viability Kit, for microscopy and quantitative analysis 1 L ,900 Mouse Anti Human CD3 delta Antibody (Clone 7D6), APC conjugate 0.5 ml MHCD ,900 MORE INFO Mouse Anti Human CD4 Antibody (Clone S3.5), Alexa Fluor 488 conjugate 0.5 ml MHCD ,300 Mouse Anti Human CD8 Antibody (Clone 3B5), PE-Cy7 conjugate 0.5 ml MHCD ,300 Mouse Anti Human CD19 Antibody (Clone SJ25-C1), Pacific Blue conjugate 0.5 ml MHCD ,300 Mouse Anti Human CD45 Antibody (Clone HI30), Pacific Orange conjugate 0.5 ml MHCD ,300 page 16 page 17

10 ?? N-PER Neuronal Protein Extraction Reagent Title 3 Number 18 Name Thermo Scientific N-PER Neuronal Protein Extraction Reagent 30 ELISA GTPase Thermo Scientific Pierce BCA Protein Assay Syn-PER Synaptic Protein Extraction Reagent 1 mg µg µg BCA Protein Assay N-PER Neuronal Protein Extraction Reagent T-PER PBS PBS 0.1% Triton X-100 Pierce BCA ProteinAssay Reagent T-PER Tissue Protein Extraction Reagent mg 10 µg µg 10 µg H S C Syn 200 mg 100 mmhepes 3 post-synaptic density protein 95 PSD95 NMDA receptor 2B subunit AMPA receptors GluR2/3/4Calcineurin Cdk5 HDAC2 -actin N-PER Neuronal Protein Extraction Reagent 10 g10 cm ml ,300 Syn-PER Synaptic Protein Extraction Reagent 10 g mm ml ,700 DNA 90 2 Paul Russell Russell DNA Paul Russell Paul Russell Russell Square Paul Russell 1 2 Molecular Biology of the Cell Paul Russell page 18 page 19

11 ネガコン一番光ってるINFORMATION 2016 / July / No.36 in the 2016 STARD10 光ってる NEXT 7? T AmpliSeq 5Quo 5 Quo! photographs: ryoichi morikawa p.02-03, p art direction & design: masami furuta opportune design science writing: momoyo matsuyama, hiroshi mizuno edition: yuko hashimoto NEXT "" 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. TaqMan is a registered trademark of Roche Molecular System, Inc. Printed in Japan. LSG008-A1606HS TEL FAX

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IonTorrentPGM_appnote_0319.indd Ion PGM Heterogeneity * 1 * 1 * 2 * 2 * 2 * 2 * 2 * 1 *1 * 2 DNA DNA DNA ArcturusXT LCM Laser Capture Microdissection LCM A Workflow for Cancer Profiling Applied Biosystems ViiA7 PCR qpcr QC Assay Ion