Eccles:cDNA Synthesis

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cDNA Synthesis:

Part 1

  • Per Reaction:
    • 1μL 250ng/μL 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).
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