Team:DTU-Denmark/Notebook/8 August 2013

From 2013.igem.org

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(Procedure)
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! component(per reaction) !! without additives !! using 5% DMSO !! using 1M Betaine
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===PCR for extraction of Nir1===
===PCR for extraction of Nir1===

Revision as of 08:06, 9 August 2013

8 August 2013

Contents

lab 208


Main purpose


Who was in the lab


Kristian, Julia, Henrike

Procedure


PCR for AMO with USER endings

primers: 17a, 17b

template: gel purified AMO extraction fragment

Program: Standard with 54C annealing temperature and 3:00 extension time


}

PCR for extraction of Nir1

Results



Gel on yesterdays PCR

  • 1kb ladder
  • Yesterdays sample 1
  • Yesterdays sample 2
  • Yesterdays sample 3
  • Yesterdays sample 4
  • Neg.
  • Neg.
  • Nir2
  • cycAX Histag
  • 1kb ladder

2013-08-08 unkown gel.jpg


Gel on ON PCR samples

  • 1kb ladder
  • Nir1
  • Nir1 5%DMSO
  • Nir1 1M Betaine
  • Nir1 3M Betaine
  • Nir2 3M Betaine U
  • Nir2 1M Betaine U
  • Nir2 5%DMSO U
  • Nir2 U
  • Neg (Nir)
  • Ref 0%DMSO
  • Ref 2%DMSO
  • Ref 5%DMSO
  • AMO U
  • AMO 2%DMSO U
  • Neg (AMO)
  • NirG 5%DMSO
  • NirG
  • 1kb ladder

2013-08-08 Nir.jpg


Gel on todays PCR samples

  • 1kb ladder
  • Nir Q5 poly
  • Nir 2%DMSO Q5 poly
  • Nir 3M Betaine Q5 poly
  • Nir 5%DMSO Q5 poly
  • Nir1 Q5 poly
  • Nir1 2%DMSO Q5 poly
  • Nir1 5%DMSO Q5 poly
  • Nir1 3M Betaine Q5 poly
  • AMO USER primers
  • AMO +DMSO USER primers
  • AMO +Betaine USER primers
  • Nir1 DMSO
  • Nir1 Betaine
  • 1kb ladder

2013-08-08 nir1 AMO user.jpg

Conclusion


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component(per reaction) without additives using 5% DMSO using 1M Betaine
1uL | 1 uL | 1uL
HF buffer
X7 polymerase
MilliQ water
template
FW primer
RV primer
DMSO
Betaine

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