EPF-Lausanne/5 August 2009

From 2009.igem.org

(Difference between revisions)
(Wet Lab)
(Wet Lab)
 
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From the transformations of yesterday, only 3/16 plates worked: the "classic method" LovTap-Term, its negative control (probably the Term plasmid religated), and one clone on the LacI-RBS-LovTap-Term!!! We suspect that the death cassette plasmid doesn't work correctly (since this hasn't been successful from the beginning), so next time we'll use other backbones for the ligation.
From the transformations of yesterday, only 3/16 plates worked: the "classic method" LovTap-Term, its negative control (probably the Term plasmid religated), and one clone on the LacI-RBS-LovTap-Term!!! We suspect that the death cassette plasmid doesn't work correctly (since this hasn't been successful from the beginning), so next time we'll use other backbones for the ligation.
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We did a colony PCR (with the iGEM primers) on 20 clones from the LovTap-Term, 5 from the negative control, and the one from the LacI-RBS-LovTap-Term, and ran the resulting fragments on an agarose gel to see whether the inserts were the expected ones. Result: only one of the 20 clones (LacI-RBS) was good.
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We did a colony PCR (with the iGEM primers) on 20 clones from the LovTap-Term, 5 from the negative control, and the one from the LacI-RBS-LovTap-Term, and ran the resulting fragments on an agarose gel to see whether the inserts were the expected ones. Result: only one of the 20 clones (LovTap-Term) was good.
We did a liquid culture of the right clone.
We did a liquid culture of the right clone.

Latest revision as of 09:40, 10 August 2009

Contents

5 August 2009





Wet Lab

From the transformations of yesterday, only 3/16 plates worked: the "classic method" LovTap-Term, its negative control (probably the Term plasmid religated), and one clone on the LacI-RBS-LovTap-Term!!! We suspect that the death cassette plasmid doesn't work correctly (since this hasn't been successful from the beginning), so next time we'll use other backbones for the ligation.

We did a colony PCR (with the iGEM primers) on 20 clones from the LovTap-Term, 5 from the negative control, and the one from the LacI-RBS-LovTap-Term, and ran the resulting fragments on an agarose gel to see whether the inserts were the expected ones. Result: only one of the 20 clones (LovTap-Term) was good.

We did a liquid culture of the right clone.

To redo the ligations for what didn't work, we did the "1,5-step PCR" again and the normal PCR with the 3 LacI-RBS we have.

People in the lab

Nath, Basile, Gab, Christian, Nicolas