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EPF-Lausanne/28 July 2008
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The gel revealed that the PCR from last Friday did not work, so we did a new PCR to check a control with test DNA and test primer, with something that worked, in two technical replicates. If it works, we will use it as positive control in the following PCRs. The PCR from the prof. worked, but ours didn't, so perhaps the dNTP concentration might be wrong, or the pipeting was done wrong. | The gel revealed that the PCR from last Friday did not work, so we did a new PCR to check a control with test DNA and test primer, with something that worked, in two technical replicates. If it works, we will use it as positive control in the following PCRs. The PCR from the prof. worked, but ours didn't, so perhaps the dNTP concentration might be wrong, or the pipeting was done wrong. | ||
Revision as of 12:45, 4 August 2008
The gel revealed that the PCR from last Friday did not work, so we did a new PCR to check a control with test DNA and test primer, with something that worked, in two technical replicates. If it works, we will use it as positive control in the following PCRs. The PCR from the prof. worked, but ours didn't, so perhaps the dNTP concentration might be wrong, or the pipeting was done wrong.
A new tentative at electroporation was done, using pUC as well as 1000 2C (something useless to us), with the same protocol as before. Our cells are probably not competent.
We also transform with heat shock normal protocol our Top10 cells with the purified stock of what we received from iGEM last week.
Finally, we incubate the last of the parts which had previously not worked.
