EPF-Lausanne/15 September 2008
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[https://2008.igem.org/wiki/index.php?title=EPF-Lausanne/22_September_2008 Next>>] | [https://2008.igem.org/wiki/index.php?title=EPF-Lausanne/22_September_2008 Next>>] | ||
+ | ==Sequencing== | ||
[[Image:epfl_gel-09-15-08.jpg|thumb|Agarose gel: Testing primers before sequencing]] | [[Image:epfl_gel-09-15-08.jpg|thumb|Agarose gel: Testing primers before sequencing]] | ||
[[Image:epfl_gel-09-15-08_2.jpg|thumb|Agarose gel: Annealing temp. reduced to 53°C for AB-3]] | [[Image:epfl_gel-09-15-08_2.jpg|thumb|Agarose gel: Annealing temp. reduced to 53°C for AB-3]] | ||
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Before sending the plasmids for sequencing we test our primers by PCR. | Before sending the plasmids for sequencing we test our primers by PCR. | ||
Latest revision as of 19:25, 28 October 2008
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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.