Eccles:cDNA Synthesis

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cDNA Synthesis:

Part 1

• Per Reaction:
  • 1μL 250ng/uL Random Primer
  • 1μL 10mM dNTPs
  • ? μL RNA (to give desired concentration)
  • MQ up to 14μL

1. 65°C for 5 minutes (Called RT1 in Shujie's folder on red PCR machine).
2. Put on ice.

Part 2

• Per Reaction (6μL):
  • 4μL 5x RT buffer
  • 1μL 0.1M DTT
  • 1μL Superscript III enzyme

1. Add to the above RNA solution.
2. 25°C for 5 minutes.
3. 50°C for 60 minutes.
4. 70°C for 15 minutes (Called RT2 in Shujie's folder on red PCR machine).