Team:Evry/Notebook

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Revision as of 14:04, 16 July 2013

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. == Labwork: == '''Friday, June 21st''' The three following E. coli strains have been prepared to be chemically competent: BL21, DH5𝝰 and TOP10. To check the quality of our work as well as their competence, each strain has been plated on LB medium with ampicillin, kanamycin or chloramphenicol. We also transformed our bacteria with a random plasmid and plated them on LB medium with ampicillin only. We incubated our petri dishes for the week end at 30°c '''Monday, June 24th''' Competents cells made on friday 21st was plated and incubated the whole week-end at 30°C. Today we analyzed results and it seems that medium had been contaminated. New competents cells will be made. '''Tuesday, June 25th''' New competent cells have been made with the same strains of bacteria. We thought that the contamination of our cells was due either to the solution of CaCl2 that we used which may had not been sterile and/or incorrect manipulations. This time, we prepared new CaCl2 solutions that we made sure to autoclave correctly. Then, same tests as we did before of contamination and competence have been made. '''Wednesday, June 26th''' The DH5𝝰 strain was not contaminated and competent. The TOP10 strain was contaminated only on the Kanamycin petri dish and competent. The BL21 strain was contaminated and not usable. '''Thurdsay, June 27th''' We designed the plasmid constructions and primers we will use. '''Tuesday, July 2nd''' We prepared the BL21 and BG1655 strains to be competent. '''Wednesday, July 3rd''' The two strains have been contaminated and are not usable. '''Thursday, July 4th''' Genomic DNA extraction of the BL21 and BG1655 strains have been done using two different methods. We received the primers we have ordered. They have been prepared to do a PCR. In order to get our genes of interest we did a PCR using as the DNA template the genomic DNA extracted from BL21 and BG1655. '''Friday, July 5th''' To ensure the amplification of our genes by PCR, we did a agarose gel electrophoresis. To do so, we first prepared a TAE 50X stock solution. By diluting it, we use a 1X TAE solution to do a 1% agarose gel. We obtain the following gel. That showed that only two of our genes have been amplificated by PCR. A new PCR will be made.