PrimeScript® RT Master Mix (Perfect Real Time)
Contents
1. http www takara bio coJjp 8 RR036A B RNA RNA RNase RNase RNA RNase RNA 1 2 1 180 60 2 0 1 DEPC 37C 12 DEPC 120 30 RNA2. PCR http wwwr takara bio coJp 7 RR036A VIl Appendix A cDNA PrimeScript RT Master Mix Perfect Real Time Human Placenta total RNA 2 pg 2 ug 20 ul 37 15 30 60 85 5 4 PCR SYBR Premix Ex Tag ll Perfect Real Time 2 ul 25 ul Human ACTB Perfect Real Time ThermalCycler Dice Real Time System 15 ss FE 30 0 0 1 i 15 min 0 999 99 0 Y 3 346 LOG X 34 52 30 min 1 000 98 9 Y 3 349 LOG X 34 46 3301 IOG00 3424 15 30 60
3. RNA 180 60 DEPC 0 1 DEPC RNA RNA RT PCR RNA RNA GTC RNA NucleoSpin RNA II 740955 10 RNAiso Plus 9108 9109 RNA total RNA RNA TE Buffer DNA total RNA
4. 20 http wwwr takara bio coJp 2 RR036A lI 1 SNA 2 PCR cDNA 2 RT PCR 3 Oligo dT Primer Random 6mers 2 PCR cDNA 4 RT PCR total RNA cDNA TE EASY Dilution for Real Time PCR IV
5. DNA DNA PCR DNA total RNA 1 DNA 2 DNase DNA 1 DNA DNA DNA 2 DNA PCR
6. SYBR Molecular Probe Inc http wwwr takara bio coJp 11 RR036A 4KaR4 Tel 077 543 6116 Fax 077 543 197 http www takara bio co jp v201109Da
7. DNA DNA mRNA PCR DNase DNA DNA http wwwr takara bio coJp 10 RR036A VII PrimeScript RT reagent Kit Perfect Real Time RR037A B PrimeScript RT reagent Kit with gDNA Eraser Perfect Real Time RR047A B SYBR Premix Ex Tag 11 Tli RNaseH Plus RR820A B SYBR Premix Ex Tag Tli RNaseH Plus RR420A B SYBR Premix Ex Tag GC Perfect Real Time RR071A B SYBRS Premix DimerEraser Perfect Real Time RR091A B Premix Ex Tag Probe qPCR RR390A B EASY Dilution for Real Time PCR 9160 Thermal Cycler Dice Real Time System
8. 1 5 X PrimeScript RT Master Mix 5 X PrimeScript RT Master Mix 2 3 Oligo dT Primer Random 6mers Gene Speific Primer http wwwr takara bio coJp 3 RR036A V RNA VII B RNA 9 1 1 gt 5 X PrimeScript RT Master Mix Perfect Real Time 2 ul 1 Xx total RNA RNase Free dH2O up to 10 wl
9. mRNA 2 DNasel http wwwr takara bio coJp 9 RR036A 2 DNase DNA NucleoSpin RNA I RNA DNase RNA total RNA Recombinant DNasel RNase free 2270A DNA DNase 1 total RNA 20 50 pg 10 Xx DNase Buffer 5 ul RNase Inhibitor 20U DNasel RNase free 2ul 10U DEPC
10. PCR 95 5 15 PCR 95C 30 3 Thermal Cycler Dice Real Time System http www takara bio coJp 5 RR036A lt Applied Biosystems 7300 7500 7500 Fast Real Time PCR System StepOnePlus Real Time PCR System gt 1 PCR 1 gt SYBR Premix Ex Tag I Tli RNaseH Plus 2X 10 gil 25 ul 1 PCR Forward Primer 10 uM 0 8 ul 2 ul 04 uM PCR Reverse Primer 10 uM 0 8 ul 2 Mul 04 uM 1 ROX Reference Dye 50 X or Dyell 50 Xx 2 0 4 ul 1 ul 1Xx RT cDNA 2 ul 4 ul 3 dHzO 6 ul 16 wl total 20 ul 4 50 4
11. 10 500 ng total RNA 2 37C 15 85 5 4 2 PCR PCR 10 http wwwr takara bio coJp 4 RR036A VI PCR SYBR Premix Ex Tag 1 Tli RNaseH Plus RR820A PCR PCR TaqMan Premix Ex Tag Probe qPCR RR390A Thermal Cycler Dice Real Time System gt 1 PCR 1 gt SYBR Premix Ex Tag 1 Tli RNaseH Plus 2X 12 5 ul 1Xx PCR Forward Primer 10 uM 1 ul 0 4 uM 1 PCR Reverse Primer 10 uM 1 ul 0 4 uM 1 RT CDNA 2 ul
12. 50 ul up 2 37 20 3 DNase A 1 2 5 ul 0 5M EDTA 80 2 2 DEPC 100 ul fup B 1 50 ul DEPC 100 ul 25 24 1 2 15 000 rpm 5 3 24 1 4 15 000 rpm 5 10 ul 3 M 250 ul 10 4C 15 000 rpm 15 70 4 15 000 rpm 5 DEPC CoO M OD WW DNA PCR
13. TP900 TP960 NucleoSpin RNAII 740955 10 50 250 NucleoSpin RNA XS 740902 10 50 250 RNAiso Plus 9108 9109 Perfect Real Time RefSeq RT PCR SYBR Premix Ex Tag TIi RNaseH Plus SYBR Premix Ex Tag Tli RNaseH Plus SYBR Green RT PCR http www takara bio coJp realtime IX 1 1999 44 189 193 2 1997 PCR Tips 94 99
14. 1 primer 0 4 uM 0 2 1 0 uM 2 ROX Reference Dye ll 50X ROX Reference Dye 50X Applied Biosystems 7500 7500 Fast Real Time PCR System ROX Reference Dyell 50X StepOnePlus 7300 ReaLTime PCR System ROX Reference Dye 50 3 20 total RNA 10 pg 100 ng cDNA template PCR 10 4 2 PCR PCR PCR Tm PCR 3 PCR lt Applied Biosystems 7300 7500 Real Time PCR System StepOnePlus gt
15. 2 dH2O 8 5 ul Total 25 ul 1 primer 0 4 uM 0 2 1 0 uM 2 total RNA 10 pg 100 ng cDNA template PCR 10 2 PCR PCR PCR Tm PCR 3 PCR PCR Hold Cycle 1 95 30 2 Step PCR Cycle 40 95 5 60 30 Dissociation Dow otcsee 7aKafRa Ex Tag HS 7ag PCR PCR
16. 3 EASY Dilution for Real Time PCR 2 1 ml 1 PrimeScript RTase RNase Inhibitor Oligo dT Primer Random 6mers dNTP Mixture 2 Mg2 total RNA cDNA TE EASY Dilution PCR PCR EASY Dilution for Real Time PCR 9160 EASY Dilution PCR 37 85 0 2 ml 1 5 ml
17. BOOeR Regip Re PCR stage1 Stage 2 Stage 3 Stage 1 Reps 1 95 30 Stage 2 PCR Reps 40 95 5 60 30 34 Stage 3 Melt Curve StepOnePlus 30 7300 31 7500 34 http www takara bio coJp 6 RR036A Applied Biosystems 7500 Fast Rea Time PCR System gt PCR Holding Stage Reps 1 95 30 Cycling Stage Number of Cycles 40 95 3 60 30 Melt Curve Stage 7aKaRa Ex 7ag H5 7ag PCR PCR PCR 95 5 15 PCR 95 30 3
18. wm RRO36A Tahafa PrimeScript RT Master Mix Perfect Real Time v201109Da 2 RT PCR RT PCR RT PCR PrimeScript RTase RNase Inhibitor Random 6mers Oligo dT Primer dNTP Mixture 5x RNA PrimeScript RTase PrimeScript RT reagent Kit Perfect Real Time RR037A B PCR cDNA cDNA SYBR Green TaqMan SYBR Premix Ex 7gg 1 Tli RNaseH Plus SYBRS Premix Premix Ex 7gg Probe qPCR PCR 1 10 Al X 200 1 5 X PrimeScript RT Master Mix Perfect Real Time 1 400 pil 2 RNase Free dH20 1 mlX2
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