製品コード CY505S) Cycleave®PCR Reaction Mix （製品コード CY505A
Contents
1. O http www takara bio co jp 10 CY505S CY505A VII Positive Control CycleavePCR Thermal Cycler Dice Real Time System CycleavePCR Starter Kit Positive Control 1 PCR 1 gt CycleavePCR Reaction Mix 2Xconc 12 5 ul 1x Positive Control Primer Mix 10 u M each 1 ul 0 2 uM Positive Control Probe FAM TAMRA 25 X conc 1 ul 1X Positive Control 1 ul dH2O 9 5 ul total 25 ul 2 Pattern 3 Step PCR 45 55 0 72 0 00 10 0620 Bata Collection r F 3 Positive Control Positive Control 4 0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 H 35 39 40 42 44 46 Cycles http www takara bio co jp 112. PCR 10 3 Step PCR Hold Cycle 1 95 10 30 3 Step PCR Cycle 45 95 C 5 55 10 A ac z 72 20 TaKaRa Ex Taq HS 7ag PCR PCR PCR 95 C 5 15 PCR 95 C 30 3 Thermal Cycler Dice Real Time System http ww
3. PCR 10 3 Step PCR Stage 2 Hold Repeat 45 times Cycle 1 95 C 10 30 Stage 1 Temp Secs Optics Off Deg Sec Temp Secs Optics NA 96 0 5 Of NA 65 0 no Off 3 Step PCR NA 72 0 10 On Cycle 45 95 C 5 55 C 10 72C 10 15 TaKaRa Ex Tag HS 7ag PCR PCR PCR 95 C 5 IS PCR 95 C 30 3 Sma
4. 1 PCR PCR DNA DNA DNA 3 1 100 PCR TaKaRa Ex Taq HS 2 RNA DNA RNase H http www takara bio co jp 2 CY505S CY505A QTO OnO TO O
5. CycleavePCR Assay Designer CycleavePCR Assay Designer 10 Tm lt HR gt http www takara bio coJp 12 CY505S CY505A IX Appendix RT PCR RT PrimeScript RT Master Mix Perfect Real Time RR036A PrimeScript RT Master Mix Perfect Real Time Oligo dT Primer Rando
6. 500 ng total RNA 2 37 C 15 2 85 5 4 2 Gene Specific Primer 42 15 PCR 50 3 PCR RT CDNA CycleavePCR 10 http www takara bio co jp 13 CY505S CY505A X Thermal Cycler Dice Real Time System TP900 TP960 Thermal Cycler Dice Real Time System Lite TP700 TP760 Cycleave human ALDH2 Typing Probe Primer Set Ver 2 CY403 PrimeScript RT Master Mix Perfect Real Time RRO36A B PrimeScript RT reagent Kit Perfect Real Time RRO37A B Cycleave PCR Assay Designer SNPs gt http www takara bio co jp realtime_snps CycleavePCR Assay Designer gt http www takara bio co jp prt primer intro htm XI SNP
7. PCR http www takara bio coJp 8 CY505S CY505A lt LightCycler gt LightCycler Roche Diagnostics 1 PCR ROX Reference Dye 1 gt CycleavePCR Reaction Mix 2Xconc 10 ul 1x PCR Forward Primer 10 uM 0 4 ul 0 2 uM PCR Reverse Primer 10 uM 0 4 ul 0 2 uM 5 uM 0 8 ul 0 2 uM 1 template lt 100ng 1 ul 2 dH2O 7 4 ul total 20 ul x1 0 2 uM 2 1 template dH20 cDNA RT template PCR 10 2 PCR Lig
8. http www takara bio co jp 9 CY505S CY505A 3 Step PCR PCR 95 10 30 95 10 30 DNA 95 1 2 2 3Step PCR 95 C 5 PCR 300 bp 95 3 10 55 62 C 10 20
9. CycleavePCR Assay Designer SNPs CycleavePCR Starter Kit CY505S CycleavePCR Reaction Mix CY505A B TaqMan PCR Premix Ex Tag Probe qPCR RR390A B SYBR Green PCR SYBR Premix Ex Tag Tli RNaseH Plus RR420S A B SYBR Premix Ex Taq II Tli RNaseH Plus RR820S A B SYBR Molecular Probes Inc TaqMan Roche Diagnostics Thermal Cycler Dice Real Time System TP900 TP960 Thermal Cycler Dice Real Time System Lite TP700 TP760 Smart Cycler System Smart Cycler Il System Cepheid Applied Biosystems 7500 7500 Fast Real Time PCR System StepOnePlus Real Time PCR System Life Technologies LightCycler Roche Diagnostics Smart Cycler Cepheid 1 CycleavePCR Reaction Mix t amp TaKaRa Ex Taq HS PCR PCR
10. PTY RNA RNase H RNA RNA RNA 1 5 RNase H 1 ll CycleavePCR Starter Kit CY505S 80 25 ul CycleavePCR Reaction Mix 2 X conc 1 1 ml ROX Reference Dye 50 X conc 2 200 ul ROX Reference Dye Il 50 X conc 2 200 ul Positive Control 3 10 ul Positive Control Primer Mix 10 uM each 3 10 ul Positive Control Probe FAM TAMRA 25 X conc 4 10 ul dH20 1 ml CycleavePCR Reaction Mix
11. Tel 077 543 6116 Fax 077 543 1977 http www takara bio co jp SII AA RNS AL v201311Da
12. 72 C 30 300 bp 30 Smart Cycler 15 Applied Biosystems 7500 Real Time PCR System 34 30 50 cycles O Ct PCR 100 0 1 uM 1 0 uM w PCR O
13. authorized instruments PCR OO OTNS VIl SNPs SNP 1 CycleavePCR Assay Designer SNPs 20 PCR 1 SNP CycleavePCR Reaction Mix 2Xconc template PCR
14. PCR PCR 7aKaRa Ex Taq HS PCR V 1 http wwwr takara bio coJp 4 CY505S CY505A VI lt Thermal Cycler Dice Real Time System gt 1 PCR ROX Reference Dye 1 gt Cycleav
15. 1 Esaki H Noda K Otsuki N Kojima A and Asai T 2004 J Microbiol Methods 58 1 131 4 2 Kosaka T Yatabe Y Endoh H and Yoshida K 2006 Clin Cancer Res 12 5764 5769 3 Yatabe Y Hida T Horio Y and Kosaka T 2006 J Mol Diagn 8 335 341 4 Suzuki Y Saito R Zaraket H and Daoat C 2010 J Clin Microbiol 48 57 63 Xl http www takara bio coJp 14 CY505S CY505A NOTICE TO PURCHASER LIMITED LICENSE L15 Hot Start PCR Licensed under U S Patent No 5 338 671
16. CY505A 400 25 ul CycleavePCR Reaction Mix 2 X conc 1 1m x5 ROX Reference Dye 50 X conc 2 200 ul ROX Reference Dye Il 50 X conc 2 200 ul 1 TaKaRa Ex Taq HS dNTP Mixture Mg2 TliRNaseH Il 2 Life Technologies PCR ROX ROX Reference Dye StepOnePlus Real Time PCR System Life Technologies ROX Reference Dye Il Applied Biosystems 7500 7500 Fast Real Time PCR System Life Technologies Thermal Cycler Dice Real Time System TP900 Smart Cycler System Smart Cycler II System Cepheid LightCycler Roche Diagnostics 3 4 http www takara bio co jp 3 CY505S CY505A Ill IV PCR
17. CY505S CY505A Vill 1 SNPs CycleavePCR Assay Designer SNPs CycleavePCR CycleavePCR Assay Designer SNPs 3 Tm gt 2
18. PCR 10 http www takara bio co jp 7 CY505S CY505A Hong Stage Cycling Stage umber of Cycles 45 lt Fnat s toOets staring Or 3 Step PCR 350 C 7 50 C Hold oo Cycle 1 95 10 30 nane roo 3 Step PCR kr Cycle 45 95 5 55 C 10 72C 20 25 StepOnePlus ral Siep 1 Stee map 2 S03 20 7500 Fast 25 TaKaRa Ex Taq HS 7ag PCR PCR PCR 95 C 5 15 PCR 95 C 30 3
19. and 5 587 287 and corresponding patents in other countries L52 Rox Reference Dye Research Field Use of this product is covered by one or more of the following US patents and corresponding patent claims outside the US 5 928 907 The purchase of this product includes a limited non transferable immunity from suit under the foregoing patent claims for using only this amount of product for the purchaser s own internal research No right under any other patent claim and no right to perform commercial services of any kind including without limitation reporting the results of purchaser s activities for a fee or other commercial consideration is conveyed expressly by implication or by estoppel This product is for research use only Further information on purchasing licenses may be obtained by contacting the Director of Licensing Applied Biosystems 850 Lincoln Centre Drive Foster City California 94404 USA M40 Thermostable RNase H This product is covered by the claims of U S Patent No 7 422 888 and its foreign counterpart patent claims M46 PCR Cycleave This product is covered by the claims of Japanese Patent No 4022522 M57 LA Technology This product is covered by the claims 6 16 outside the U S corresponding to the expired U S Patent No 5 436 149 http www takara bio co jp 15 CY505S CY505A TafdR4
20. ePCR Reaction Mix 2Xconc 12 5 ul 1X PCR Forward Primer 10 uM 0 5 ul 0 2 uM PCR Reverse Primer 10 uM 0 5 ul 0 2 uM 5 uM 1 ul 0 2 uM 1 template lt 100 ng 1 ul 2 dH2O 9 5 ul total 25 ul 1 0 2 uM Thermal Cycler Dice Real Time System Result Analysis Data Amplification Plots Fluorescence Raw 10 000 5 000 30 000 2 1 pl template dH20 cDNA RT template PCR 10 2 PCR PCR 3 Step PCR
21. htCycler PCR 3 Step PCR PCR 10 3 Step PCR Hold Cycle 1 95 10 30 3 Step PCR Cycle 45 95 5 55 10 72 10 15 TaKaRa Ex Tag HS Tag PCR PCR PCR 95 C 5 15 PCR 95 C 30 3 LightCycler
22. m 6mers O m mRNA cDNA Gene Specific Primer PrimeScript RT Reagent Kit Perfect Real Time RR037A 1 RNA o RNA lt 1 gt RRO36A 5 XPrimeScript RT Master Mix for Real Time 2 ul 1x total RNA RNase Free dH2O up to 10 ul lt 1 gt RRO37A 5 XPrimeScript Buffer for Real Time 2 ul 1x PrimeScript RT Enzyme Mix 0 5 ul Oligo dT Primer 50 uM 1 0 5 ul 25 pmol total RNA RNase Free dH2O up to 10 ul 1 Random Primer Gene Specific Primer Random 6mers 100 uM 0 5 ul 50 pmol Gene Specific Primer 2 uM 0 5 ul 1 pmol 10 ul
23. rt Cycler System http www takara bio co jp 6 CY505S CY505A lt Applied Biosystems 7500 Fast Real Time PCR System StepOnePlus Real Time PCR System gt 1 PCR 1 gt CycleavePCR Reaction Mix 2Xconc 10 ul 1x PCR Forward Primer 10 uM 0 4 ul 0 2 uM PCR Reverse Primer 10 uM 0 4 ul 0 2 uM 5 uM 0 8 ul 0 2 uM 1 ROX Reference Dye 50Xconc or Dye ll 50Xconc 2 0 4 ul 1x template lt 100 ng 1 0 ul 3 dH2O 7 0 ul total 20 ul x1 0 2 uM 2 ROX Reference Dye II 50X ROX Reference Dye 50X StepOnePlus ROX Reference Dye 50X 7500 Real Time PCR System 7500 Fast Real Time PCR System ROX Reference Dye Il 50x ROX ROX Reference D
24. w takara bio co jp 5 CY505S CY505A lt Smart Cycler II System gt 1 PCR ROX Reference Dye 1 gt CycleavePCR Reaction Mix 2Xconc 12 5 ul 1x PCR Forward Primer 10 uM 0 5 ul 0 2 uM PCR Reverse Primer 10 uM 0 5 ul 0 2 uM 5 uM 1 ul 0 2 uM 1 template lt 100 ng 1 ul 2 dH2O 9 5 ul total 25 ul x1 0 2 uM Smart Cycler 300 500 2 1 template dH20 cDNA RT template PCR 10 2 Smart Cycler Smart Cycler PCR 3 Step PCR
25. waar CY505S waar CY505A IdfaR4 Cycleave PCR Starter Kit CY505S Cycleave PCR Reaction Mix CY505A v201311Da CycleavePCR Reaction Mix PCR 2 x 7ag in PCR AER TaKaRa Ex Taq Hs PCR 1 CycleavePCR Starter Kit CycleavePCR Reaction Mix Positive Control CycleavePCR Reaction Mix SNP 1
26. ye 3 1 pl template dH20 cDNA RT template PCR 10 2 PCR Experiment Properties Quantification Standard TaqMan Reagents Others Others Include Melt Curve Plate Set Up Define Target Reporter Quencher Plate Layout Passive Reference ROX ROX ROX Reference Dye Passive Reference none Quencher Eclipse Quencher none PCR 3 Step PCR
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