Team:DTU-Denmark/Notebook/6 September 2013

From 2013.igem.org

(Difference between revisions)
(Results)
(Gel 2)
Line 64: Line 64:
* HAO User on 59C, 3% DMSO
* HAO User on 59C, 3% DMSO
* HAO User on 59C, 5% DMSO
* HAO User on 59C, 5% DMSO
 +
* 2-log ladder
 +
 +
===Gel 3===
 +
 +
* 2-log ladder
* HAO User on touchdown, 3% DMSO
* HAO User on touchdown, 3% DMSO
* HAO User on touchdown, 5% DMSO
* HAO User on touchdown, 5% DMSO

Revision as of 15:49, 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


Gel 1

Gel of yesterday's PCR was empty (we lost the picture though...)

PCR was redone.

Gel 2

For the PCR results of today.

  • 2-log ladder
  • HAO User on 59C, 3% DMSO
  • HAO User on 59C, 5% DMSO
  • 2-log ladder

Gel 3

  • 2-log ladder
  • HAO User on touchdown, 3% DMSO
  • HAO User on touchdown, 5% DMSO
  • 2-log ladder

Conclusion



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