SBB13Ntbk - Mar12

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Regular Zymo Cleanup

  1. Add 180 uL of Zymo ADB buffer (brown bottle) to a 33uL or 50uL reaction.
  2. Transfer into the Zymo column (small clear guys)
  3. spin through (top speed, 30 sec), discard waste.
  4. Add 200 uL of Zymo Wash Buffer (which is basically 70% ethanol)
  5. spin through, discard waste.
  6. Add 200 uL of Zymo Wash Buffer
  7. spin through, discard waste.
  8. spin for 90 seconds, full speed to dry.
  9. elute with water into a fresh Eppendorf tube, use the same volume of water as the volume of the original reaction

Amplification

  1. 1 ul each outer oligo (10 uM)
  2. 1 ul purified pca product from March 8
  3. .5 ul phusion
  4. 10 ul 5x phusion buffer
  5. 5 ul 2mM dNTPs
  6. 32.5 ul H2O
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