One Step PrimeScript® RT-PCR Kit (Perfect Real Time)
Contents
1. RNase RNA 1 2 1 180 60 2 0 1 DEPC 37 12 DEPC 120 30 RNA RNA 180 60 DEPC 0 1 DEPC RN2. 100 Hot Start PCR TaKaRa Ex Taq HS 2 RT PCR RNA PCR RNA cDNA PCR RNA RT PCR RNA One Step RT PCR One Step RT PCR PCR Specific Primer Reverse cDNA Specific primer Forward Reverse PCR Random Primer Oligo dT Primer http wwwr takara bio coJp 2 RR064A Specific Primer Reverse Synthesis of 1st strand cDNA with PrimeScript RTase 3
3. 1 gt 2 x One Step RT PCR Buffer III 10 ul 1x TaKaRa Ex Tag HS 5 U ul 0 4 ul PrimeScript RT enzyme Mix Il 0 4 ul PCR Forward Primer 10 u M 0 4 ul 0 2 uM PCR Reverse Primer 10 u M 0 4 ul 0 2 uM TaqMan 0 8 ul 2 total RNA 2 ul 3 RNase Free dH2O 5 6 ul Total 20 ul 1 0 2 UM 0 1 1 0 UM x2 PCR 3 total RNA 10 pg 100 ng template 2 PCR LightCycler PCR Sa 12 PCR R Ocie f
4. RT PCR Mouse Actb TaqMan Gene Expression Assays Life Technologies TaqMan 2 PCR Crossing Point CP Ww x 5 gt ta E RSq 0 999 E Eff 96 4 s fa s 3 5 te 2nd Derivative Maximum SDM Dvaras Ord wwwrel BBe e02 8 38H 3 total RNA 1 pg 100 ng http www takara bio coJp 13 RR064A VIII Appendix RNA RNA cDNA PCR cDNA RNA RNase RNase RNA
5. SYBR Molecular Probe Inc TaqMan Roche Diagnostics Premix Ex Taq http wwwr takara bio coJp 15 RR064A NOTICE TO PURCHASER LIMITED LICENSE P7 PCR Notice Alicense to perform the patented 5 Nuclease Process for research is obtained by the purchase of i both Authorized 5 Nuclease Core Kit and Licensed Probe ii a Licensed 5 Nuclease Kit or iii license rights from Applied Biosystems This product is an Authorized 5 Nuclease Core Kit Use of this product is covered by one or more of the following US patents and corresponding patent claims outside the US 5 789 224 5 618 711 6 127 155 5 677 152 and 5 773 258 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 Separate purchase of a Licensed Probe would convey rights under the applicable claims of US Patents Nos 5 538 848 5 723 591 5 876 930 6 030 787 6 258 569 and 5 804 375 claims 1 12 only and correspondi
6. 95 10 3 http www takara bio co jp 11 RR064A lt PCR gt PCR amn PCR 95 C 3 5 OFF 300 bp 95 3 5 56 64 20 30 oy 56 64 C OME 31 34 Life Technologies 30 7300 31 7500 34 http wwwr takara bio coJp 12 RR064A Vil Mouse Actb Thermal Cycler Dice Real Time System 1 Mouse Liver total RNA 1 pg 100 ng 1
7. 5 http wwwr takara bio coJp 10 RR064A 2 PCR 12 PCR Stage 1 Stage 2 Stage 3 Reps 1 Reps 1 Reps Stage 1 2 Reps 1 42 5 95 C 10 Stage 3 PCR Reps 40 95 C 5 60 31 or 34 7300 31 7500 34 TaKaRa Ex Taq Hs Taq PCR PCR PCR 95 C 5 15 PCR
8. 2 PCR Hot Start TaKaRa Ex Tag HS PCR 2 V i 10 Master Mix RNase Free dH2O Master Mix TaKaRa Ex Taq HS PrimeScript RT enzyme Mix Il 50
9. 20 20 2 X One Step RT PCR Buffer Ill Vortex Random Primer Oligo dT Primer http wwwr takara bio coJp 5 RR064A VI RNA VIII RNA lt Thermal Cycler Dice Real Time System Lite gt 1 lt 1 gt 2 X One Step RT PCR Buffer III 12 5 ul 1x TaKaRa Ex Tag HS 5 U ul 0 5 ul PrimeScript RT enzyme Mix Il 0 5 ul PCR Forward Primer 10 u M 0 5 ul 0 2 uM
10. 1 gt 2 x One Step RT PCR Buffer III 10 ul 25 ul 1x TaKaRa Ex Taq HS 5 U ul 0 4 ul 1 Al PrimeScript RT enzyme Mix II 0 4 ul 1 ul PCR Forward Primer 10 u M 0 4 ul 1 ul 0 2 uM 1 PCR Reverse Primer 10 u M 0 4 ul 1 ul 0 2 uM 1 TaqMan 0 8 ul 2 ul 2 ROX Reference Dye or Dye II 50X 3 0 4 ul 1 ul total RNA 2 ul 4 ul 4 RNase Free dH2O 5 2 ul 14 ul Total 20 ul gt 50 pul 5 1 0 2 UM 0 1 1 0 UM x2 PCR 3 ROX Reference Dye Il 50X ROX Reference Dye 50X Applied Biosystems 7500 Real Time PCR System ROX Reference Dye Il 50x Applied Biosystems 7300 Real Time PCR System ROX Reference Dye 50 X 4 50 total RNA 20 pg 200 ng template
11. PCR Reverse Primer 10 u M 0 5 ul 0 2 uM 1 TaqMan 1 ul 2 total RNA 2 ul 3 RNase Free dH2O 75 ul Total 25 ul K 1 0 2 UM 0 1 1 0 UM x2 PCR Thermal Cycler Dice Real Time System 0 1 0 5 uM 3 totalRNA10 pg 100 ng template http wwwr takara bio coJp 6 RR064A 2 Thermal Cycler Dice Real Time System PCR 12 PCR
12. Pattern 2 Step PCR Segrnent Pattern 1 Hold 42 5 95 C 10 Pattern 2 PCR Temperature deg Cycle 40 Hold Time ramess 95 C 5 Data Collection 60 C 30 TaKaRa Ex Taq HS g PCR PCR PCR 95 C 5 15 PCR 95 10 3 http www takara bio co jp 7 RR064A lt Smart Cycler II System gt 1 lt 1 gt 2 X One Step RT PCR Buffer III 12 5 ul 1x TaKaRa Ex Tag HS 5 U ul 0 5 ul PrimeScript RT enzyme
13. 9 5 Synthesis of 2nd strand cDNA with TaKaRa Ex Taq HS Specific Primer Forward 1 RT PCR http www takara bio co jp 3 RR064A 3 TaqMan 5 FAM 3 TAMRA KE TaqMan DNA 7aqg DNA 5 3 exonuclease TaqMan 1 oe 2 1 1 em a 3 K ll 100 50 ul 1 2 Xx One Step RT PCR Bufferlll 1 840 ul x3 2 TaKaRa Ex Tag HS 5 U ul 100 ul 3 PrimeScript RT e
14. A RNA RT PCR RNA RNA RNA total RNA NucleoSpin RNA 740955 10 50 250 AGPC RNAiso Plus 9108 9109 http wwwr takara bio coJp 14 RR064A IX Premix Ex Taq Probe qPCR RR390A B One Step SYBR PrimeScript RT PCR Kit Perfect Real Time RRO66A B One Step SYBR PrimeScript RT PCR Kit II Perfect Real Time RRO86A B One Step SYBR PrimeScript PLUS RT PCR Kit Perfect Real Time RR096A B SYBR Premix Ex Taq Tli RNaseH Plus RR820S A B SYBR Premix Ex Taq Tli RNaseH Plus RR420S A B SYBR Premix DimerEraser Perfect Real Time RR091A B Thermal Cycler Dice Real Time System TP900 TP960 Thermal Cycler Dice R
15. Mix Il 0 5 ul PCR Forward Primer 10 u M 0 5 ul 0 2 uM 1 PCR Reverse Primer 10 u M 0 5 ul 0 2 uM 1 TaqMan 1 ul 2 total RNA 2 ul 3 RNase Free dH20 75 ul Total 25 ul 1 0 2 UM 0 1 1 0 UM 2 PCR Smart Cycler II System 0 1 0 5 UM 3 total RNA 10 pg 100 ng template 2 Smart Cycler Smart Cycler PCR 12 PCR Stage 1 Stage 2 Sta el 3 Repeat 1 times Re
16. Program D Dain Cyciws 40 9 Targets Stage 1 ne 42 5 20C Tez 95 C 10 20 9 1 Stage 2 PCR 95 5 20 60 20 20 C Y 40 Stage2 PCR TaKaRa Ex Taq HS g PCR PCR PCR 95 C 5 15 PCR 95 10 3 LightCycler http www takara bio co jp 9 RR064A lt Applied Biosystems 7300 7500 Real Time PCR System gt 1 lt
17. chnology This product is covered by the claims 6 16 outside the U S corresponding to the expired U S Patent No 5 436 149 TafdR4 Tel 077 543 6116 Fax 077 543 1977 http www takara bio co jp SII AA RNS AL v201311Da
18. e Technologies LightCycler Roche Diagnostics TaqMan Roche Diagnostics The 9 nuclease process is covered by patents owned by Roche Diagnostics Inc and F Hoffmann La Roche Ltd Purchase of the product does not provide a license to use this patented technology SYBR Green 1 RT PCR One Step SYBR PrimeScript PLUS RT PCR Kit Perfect Real Time RR096A B One Step SYBR PrimeScript RT PCR Kit Perfect Real Time RR066A B One Step SYBR PrimeScript RT PCR Kit II Perfect Real Time RR086A B One Step PrimeScript RT PCR Kit Perfect Real Time PrimeScript RTase RNA cDNA AME TaKaRa Ex Tag HS PCR 1 PCR TaqaMan 1 PCR PCR DNA DNA DNA 3 1
19. eal Time System Lite TP700 TP760 Smart Cycler Il System SC200N SC210N NucleoSpin RNA 740955 10 50 250 RNAiso Plus 9108 9109 X 1 2005 3 4 PCR 120 126 2 2008 PCR 3 1 RT PCR 39 43 Xl PrimeScript TaKaRa Ex Taq Thermal Cycler Dice DimerEraser
20. ng claims outside the United States 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 Diagnostic uses under Roche patents require a separate license from Roche Further information on purchasing licenses may be obtained from the Director of Licensing Applied Biosystems 850 Lincoln Centre Drive Foster City California 94404 USA 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 M57 LA Te
21. nzyme Mix II 100 ul 4 RNase Free dH20 1 25 ml X2 5 ROX Reference Dye 50Xconc 2 100 ul 6 ROX Reference Dye Il 50 conc 2 100 ul 1 dNTP Mixture Mo 2 Life Technologies PCR ROX Reference Dye Applied Biosystems 7300 Real Time PCR System Life Technologies ROX Reference Dye Il Applied Biosystems 7500 Real Time PCR System Life Technologies Thermal Cycler Dice Real Time System Thermal Cycler Dice Real Time System Lite e Smart Cycler System Cepheid 1 LightCycler Roche Diagnostics 1 http www takara bio co jp 4 RR064A ERED ee NS PCR authorized instruments PCR TaqMan licensed probe Ill 20 IV 1 One Step RT PCR ICKY RNA RNA
22. peat 40 times i ol d 2 Temperature Cycle v 2 Temperature Cycle Si 42 C 5 DegiSec Temp Secs Optics DegiSec Temp Secs Optics g A NA 420 300 Off NA 95 0 5 ff 95 C 10 NA 95 0 10 lof NA 60 0 20 lon Stage 2 PCR Repeat 40 95 C 5 Advance to Next Stage _ Advance to Next Stage 60 C 20 TaKaRa Ex Taq HS g PCR PCR PCR 95 C 5 15 PCR 95 10 3 Smart Cycler System http www takara bio co jp 8 RR064A lt LightCycler gt
23. ws r RRO64A IdfaR4 One Step PrimeScript RT PCR Kit Perfect Real Time v201311Da One Step PrimeScript RT PCR Kit Perfect Real Time TaqMan 1 RT PCR 2 RT PCR 1 PCR RNA RNA cDNA PrimeScript RTase PCR TakaRa Ex Taq HS 1 RT PCR 1 RT PCR x1 2 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 7300 7500 Real Time PCR System Lif
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