EPF-Lausanne/15 September 2008

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[[Image:epfl_gel-09-15-08.jpg|thumb|Agarose gel: Testing primers before sequencing]]
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[[Image:epfl_gel-09-15-08_2.jpg|thumb|Agarose gel: Annealing temp. reduced to 53°C for AB-3]]
==Sequencing==
==Sequencing==
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'''Results:'''
'''Results:'''
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[[Image:epfl_gel-09-15-08.jpg|thumb|Agarose gel: Testing primers before sequencing]]
 
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[[Image:epfl_gel-09-15-08_2.jpg|thumb|Agarose gel: Annealing temp. reduced to 53°C for AB-3]]
 
All results were as expected except for AB lane 3, i.e. AB2seqFW, LacImRV
All results were as expected except for AB lane 3, i.e. AB2seqFW, LacImRV
A new PCR is done for these primers, where the annealing temperature is reduced to 53°C.
A new PCR is done for these primers, where the annealing temperature is reduced to 53°C.
This turns out to work.
This turns out to work.

Revision as of 19:21, 28 October 2008

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Agarose gel: Testing primers before sequencing
Agarose gel: Annealing temp. reduced to 53°C for AB-3

Sequencing

Before sending the plasmids for sequencing we test our primers by PCR.

PCR on AB with:

  • pSB1A2 FW, I1466RV
  • AB1seqFW, I1466RV
  • AB2seqFW,LacImRV
  • R0071FW,pSB1A2RV

PCR on CD with:

  • pSB1A2FW, C0040RV
  • CD1FW, C0040RV
  • R0040FW, BB3
  • R0040FW, pSB1A2RV

Results:

All results were as expected except for AB lane 3, i.e. AB2seqFW, LacImRV

A new PCR is done for these primers, where the annealing temperature is reduced to 53°C. This turns out to work.