Team:DTU-Denmark/Notebook/4 August 2013

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4 August 2013

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Contents

Lab 208


Main purpose


Amplify the two Nir fragments and prepare for PCR with USER primers.

Who was in the lab


Kristian

Procedure


PCR for amplification of Nir fragments

Normal procedure for PCR on both fragments all reactions where done as duplicates:

  • Nir1 HF buffer
  • Nir1 GC buffer
  • Nir2 HF buffer
  • Nir2 GC buffer

Both Nir1 reactions where done with primer pair 41 on 68C annealing and 2:00 min extension. Both Nir2 reactions where done with primer pair 42 on 65C annealing and 2:30 min extension.


PCR on Nir1 with USER-primers

Samples where done i duplicates and with both HF- and GC-buffer. Nir1 as template and primer pair 39. 10 rounds of 60C annealing followed up by 25 rounds of 66C both had 5:00 min extension time.

Results


See gel on tomorrow's page.


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