"
EPF-Lausanne/5 August 2009
From 2009.igem.org
(New page: {{EPF-Lausanne09}} <div CLASS="epfltrick">__TOC__ </div><div CLASS="epfl09"> <html> <body> <form action="input_button.htm"> <p align="right"> <input type="button" name="lien" value="04 Au...) |
(→Wet Lab) |
||
| (3 intermediate revisions not shown) | |||
| Line 27: | Line 27: | ||
==Wet Lab== | ==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== | ==People in the lab== | ||
| - | Nath, Basile, Gab, Christian | + | Nath, Basile, Gab, Christian, Nicolas |
Latest revision as of 09:40, 10 August 2009
Contents |
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
