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From 2011.igem.org

You should make use of the calendar feature on the wiki and start a lab notebook. This may be looked at by the judges to see how your work progressed throughout the summer. It is a very useful organizational tool as well.

Note: Protocols were generally similar to those found on Open Wetware's [http://www.openwetware.org/wiki/French_Lab pages] dedicated to Chris French's Lab.

June 1 Began work to add the Plac-LacZ sequence to the pSB1C3 vector. PCR was used to introduce the B restriction site (so the final thing becomes E-X-B-Plac-LacZ-S-P.

June 2 PCR seemed successful (though 2 bands were seen on the gel, 1 was the right size). We already have pSB1C3-RFP so we cut out the RFP (using restriction enzymes XbaI and PstI) and inserted the new sequence.

June 3 After transformation of competent cells we plated onto an Xgal- and IPTG- containing media.

At this point we also prepared about 15 plates of agar, with chloramphenicol (which the pSB1C3 vector provides resistance to) as well as Xgal and IPTG for later blue/white screening.

June 6 Colonies that have successfully lost RFP and gained LacZ should be blue. We found several such colonies. Six were selected, replated and grown for 6 hours on the media mentioned above. Some cells were then transferred to bottles of media for further growth.

June 7 Plasmid DNA from the bottled cultures was purified (by the "miniprep" protocol). We digested it with EcoRI to linearise it, and then ran it on a gel. All six cultures showed a band of the correct size (about 2600 b.p.) but we will send some DNA for sequencing.

This was the old Navbox for Edinburgh; now it's obsolete...