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
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 19611986 3 1989 ADA 1994 T 2001 7 2002 3 2012 4 page 02 page 03
Ion AmpliSeq Panel User's Voice NGS DNA Ion AmpliSeq 15525 10 10% BRCA1/2 BRCA1/2 25 155 BRCA 3 25610 AmpliSeq Ion AmpliSeq Ion AmpliSeq Designer PCR BRCA1/2 25 97.6% 610 Ion Proton 15511 BRCA1/2 BRCA1/2 4 10 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): 459 466 Ion AmpliSeq Panel PCR Ion Torrent 6,144 / 1 5 Mb DNA10ng / 1 FFPE 384 1 Ion AmpliSeq DNA www.ampliseq.com 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 50 207 Comprehensive Cancer Panel 4,465 COSMIC Mutations AR, NFKB2, SOX2 409 15,992 RNA Cancer Panel KRAS, BRAF, EGFR 50 50 RNA Apoptosis Panel c-myc, TP53, FAS, BAX 267 267 TP53 Panel TP53 1 24 BRCA1 and BRCA2 Panel BRCA1, BRCA2 2 167 Colon & Lung Cancer Research Panel v2 NRAS, PTEN, PIK3CA 22 92 AML Research Panel DNMT3A, RUNX1 19 264 RNA Fusion Lung Cancer Research Panel ALK, RET, ROS1,NTRK 70 85 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 v2 8 4475346 28,000 Ion AmpliSeq Comprehensive Cancer Panel 8 4477685 108,000 Ion AmpliSeq Inherited Disease Panel 8 4477686 82,000 Ion AmpliSeq Custom Panel 7503,000 1 750 page 04 page 05
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 1 3712 SNP 1 NanoDropQubit NanoDrop One Acclaro Qubit 3.0 Fluorometer FFPE DNA Qubit 0.4ng / µl 1 Qubit 1 1 2 NanoDrop One NanoDrop One 1 A30221 1,750,000 NanoDrop One C 1 A30222 2,050,000 Qubit 3.0 Fluorometer 1 Q33216 201,000 Qubit 3.0 Quantitation Starter Kit 1 Q33217 221,100 Qubit 3.0 NGS Starter Kit 1 Q33218 216,100 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 www.thermofisher. 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 100 12351-010 17,000 Platinum SuperFi Green DNA Polymerase 100 12357-010 17,000 Platinum SuperFi PCR Master Mix 100 12358-010 43,500 Platinum SuperFi Green PCR Master Mix 100 12359-010 43,500 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
ADS! User's Voice! ADS,TaqMan Gex A29494 300,000 StepOne, StepOnePlus, 7000, 7300,7500, ADS,Custom TaqMan GEx A29496 380,000 7500Fast, 7900HT, ViiA7, ADS,Multiplex 2Dye TaqMan Gex A29498 380,000 QuantStudio 3 / 5 / 6 / 7 / 12K ADS Sequencing A29504 350,000 3130 / 3130xl, 3500 / 3500xL,3730 / 3730xl NEW! ADS,TaqMan GEx with Training A29495 420,000 StepOne, StepOnePlus, 7000, 7300,7500, ADS,Custom TaqMan GEx with Training A29497 500,000 7500Fast, 7900HT, ViiA7, ADS,Multiplex 2Dye TaqMan GEx with Training A29499 500,000 QuantStudio 3 / 5 / 6 / 7 / 12K ADS,Multiplex 3Dye TaqMan GEx with Training A29501 1,800,000 AADS,Multiplex 4Dye TaqMan GEx with Training A29503 1,800,000 ADS Sequencing with Training A29505 470,000 3130 / 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 12 4383547 11,500 450 4332077 65,900 page 08 page 09
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 93 97410 Essential 8 Flex Medium 500 ml A2858501 31,000 Vitronectin Truncated Recombinant Human VTN-N 1 ml A14700 6,000 TaqMan hpsc Scorecard Kit, Fast 96-well plate 2 1 A15871 96,000 TaqMan hpsc Scorecard Panel, Fast 96-well 2 1 A15876 70,500 page 10 page 11
! 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 CMAX00001 10,800 GeneArt Platinum Cas9 nuclease Cas9 10 µg 1µg/µL B25642 43,000 25 µg 1µg/µL B25640 85,000 GeneArt Genomic Cleavage Detection Kit 20 A24372 48,100 page 12 page 13
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 24,000 20 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-3 1 2 KOWT Click-iT EdU Colorimetric IHC Detection Kit 5-ethynyl-2'-deoxyuridine EdU DNA 1 LC-3 1 2 LC-3 WT KO1 1200 24000 20 1 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 11,2002024,000 CellInsjght CX7 20 3 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 C10644 87,800 Click-iT TUNEL Colorimetric IHC Detection Kit 1 50 C10625 97,000 page 14 page 15
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 4 2 1 100 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 2 1 4 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) 1:1 15 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 A24858 19,950,000 EVOS Onstage Incubator 1 AMC1000 1,750,000 LIVE/DEAD BacLight Bacterial Viability Kit, for microscopy and quantitative analysis 1 L7012 87,900 Mouse Anti Human CD3 delta Antibody (Clone 7D6), APC conjugate 0.5 ml MHCD0305 56,900 MORE INFO www.thermofisher.com/evosflauto2 Mouse Anti Human CD4 Antibody (Clone S3.5), Alexa Fluor 488 conjugate 0.5 ml MHCD0420 60,300 Mouse Anti Human CD8 Antibody (Clone 3B5), PE-Cy7 conjugate 0.5 ml MHCD0812 60,300 Mouse Anti Human CD19 Antibody (Clone SJ25-C1), Pacific Blue conjugate 0.5 ml MHCD1928 75,300 Mouse Anti Human CD45 Antibody (Clone HI30), Pacific Orange conjugate 0.5 ml MHCD4530 90,300 page 16 page 17
?? 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 70-90 µg10 6 300 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 78510 1 1 mg 10 µg10 6 4 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 200 100 ml 87792 38,300 Syn-PER Synaptic Protein Extraction Reagent 10 g500 35 mm500 100 ml 87793 43,700 DNA 90 2 Paul Russell Russell DNA 3 1996 Paul Russell 2 2013 3 Paul Russell Russell Square Paul Russell 1 2 Molecular Biology of the Cell 2009 2-3 1 2 7 12 199619961999 Paul Russell 9 20092013 page 18 page 19
ネガコン一番光ってるINFORMATION 2016 / July / No.36 in the 2016 STARD10 光ってる 10 50 100 2016 7 11 7 31 www.thermofisher.com/jp-senryu 2016 8 NEXT 7? T AmpliSeq 5Quo www.surveymonkey.com/s/next36 5 Quo! photographs: ryoichi morikawa p.02-03, p.10-11 art direction & design: masami furuta opportune design science writing: momoyo matsuyama, hiroshi mizuno edition: yuko hashimoto NEXT www.thermofisher.com/next 2016 7 "" www.thermofisher.com/tc For Research Use Only. Not for use in diagnostic procedures. 2016 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 108-0023 4-2-8 TEL.03 6832 9300 FAX.03 6832 9580 www.thermofisher.com