EPF-Lausanne/15 September 2008

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==Sequencing==
==Sequencing==
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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]]
Before sending the plasmids for sequencing we test our primers by PCR.
Before sending the plasmids for sequencing we test our primers by PCR.
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'''Results:'''
'''Results:'''
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[[epfl_gel-09-15-08.jpg]]
 
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

Latest revision as of 19:25, 28 October 2008

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Sequencing

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

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.