Eccles:cDNA Synthesis
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Back to Eccles Lab > DGG Protocols
cDNA Synthesis:
Part 1
- Per Reaction:
- 1μL 250ng/μL Random Primer
- 1μL 10mM dNTPs
- ? μL RNA (to give desired concentration)
- MQ up to 14μL
- 65°C for 5 minutes (Called RT1 in Shujie's folder on red PCR machine).
- Put on ice.
Part 2
- Per Reaction (6μL):
- 4μL 5x RT buffer
- 1μL 0.1M DTT
- 1μL Superscript III enzyme
- Add to the above RNA solution.
- 25°C for 5 minutes.
- 50°C for 60 minutes.
- 70°C for 15 minutes (Called RT2 in Shujie's folder on red PCR machine).