Team:DTU-Denmark/Notebook/6 September 2013

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(Procedure)
(Repeat PCR from yesterday with same conditions and programs)
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===Repeat PCR from yesterday with same conditions and programs===
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===Repeat HAO USER PCR from yesterday with same conditions and programs===
Template: HAO extraction fragment -isolated from ''Nitrosomonas europea''- (purified from PCR run on 04.09)
Template: HAO extraction fragment -isolated from ''Nitrosomonas europea''- (purified from PCR run on 04.09)

Revision as of 11:32, 6 September 2013

6 September 2013

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Contents

Lab 208


Main purpose


  • Add USER endings to HAO gene extracted from Nitrosomonas europea

Who was in the lab



Procedure


Repeat HAO USER PCR from yesterday with same conditions and programs

Template: HAO extraction fragment -isolated from Nitrosomonas europea- (purified from PCR run on 04.09)

Primers: 18a & 18b

Two versions, one with 5% DMSO and one with 3% DMSO and two programs: Touchdown (62C -> 55) and a single annealing temperature of 59C.

PCR reaction master mix

compound volume (uL)
dNTPs 1
HF 10
polymerase x7 0.5
FW 3
RV 3
template 1
MQ 29
DMSO 1.5
  • Add 49 ul of the master mix, and
  • Add 1 ul DMSO (for the 5%) or 1 ul of MQ (for the 3%)

Results


Conclusion



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