Team:Cambridge-JIC/Notebook/CL W3 Thursday
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1- checked restriction sites in UnaG- No Illegals hiding away! | 1- checked restriction sites in UnaG- No Illegals hiding away! | ||
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I have spent the morning talking with other team mwmbers on the theme of constructs - we managed to finalise a list of constructs we want to transform Marchantia with in the first round. The afternoon was dedicated to establishong Agrobacterium overnight cultures for Friday - two transformants (GFPLti and RFPLti) will be used for test Marchantia transformation, whereas another culture of non transformed bacteria will be used for creating a stock of electrocompetent cells. The primers for chromoprotein genes have arrived from Sigma, and I have created stock and working dilutions.</p> | I have spent the morning talking with other team mwmbers on the theme of constructs - we managed to finalise a list of constructs we want to transform Marchantia with in the first round. The afternoon was dedicated to establishong Agrobacterium overnight cultures for Friday - two transformants (GFPLti and RFPLti) will be used for test Marchantia transformation, whereas another culture of non transformed bacteria will be used for creating a stock of electrocompetent cells. The primers for chromoprotein genes have arrived from Sigma, and I have created stock and working dilutions.</p> | ||
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Latest revision as of 11:14, 24 July 2014
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Hugh
Work on the wiki; listened to Gos Micklem discuss good lab technique
Trang-Anh
Did a recap of which constructs and controls are to be made and which parts to get synthetised from DNA 2.0, Smolke, Japan etc. Listened to Gos's talk on lab technique. Tried to look at the Marchantia genome files from Bernardo...
Angelina
Contributed to the morning team construct planning. Learned from Barnado about the Geneious GUI,the currently annotated datasets of the marchantia genome and mRNA transcripts and the predicted ORFs. Also noted tips on how to look for promoters. Background reading.
Salil
Played around with Fernan's construct - getting nowhere. Realised there may be a way in. Collected DNA from Bernardo and loaded CC E-coli with it, for Guy to finish off a bit later. Intent upon ordering some primers tomorrow.
edit, 0106: just realised it may be possible to hack together two plasmids to build an almost full circuit.. excited to pitch plan to someone (a bit later today) with more experience in restriction/ligation cloning...
Ginny
1- checked restriction sites in UnaG- No Illegals hiding away!
2- equivocated most of the day whether to codn optimise not to codon optimise with each supervisor saying something different...conclusion: ummmmm
3- located source of 'cheap' bilirubin (unconjugated)
4- designed initial primers for UnaG constructs which are now obsolete as UnaG optimized sequence has changed...
4.bis: got confused with bioinforatics software with Angelina and Bernado- those 5' UTRs!!
5- made plans for tomorrow :P
(including catching up on the NewScientist and buying more more coffee- where is it all going?!)
Guy
I was in London for most of the day where by chance I met a farmer with all the knowledge and experience we could ever want about what would be useful to sense in agricultural soil. The outcome was that we'll be searching for phosphate inducible promoters, not nitrate ones, because nitrate tests are of less use given how easily washed away nitrates are. In the evening along with Salil I transformed and plated E. coli to amplify our chromoprotein genes, the transformant colonies should be visible in the morning.
Miha
I have spent the morning talking with other team mwmbers on the theme of constructs - we managed to finalise a list of constructs we want to transform Marchantia with in the first round. The afternoon was dedicated to establishong Agrobacterium overnight cultures for Friday - two transformants (GFPLti and RFPLti) will be used for test Marchantia transformation, whereas another culture of non transformed bacteria will be used for creating a stock of electrocompetent cells. The primers for chromoprotein genes have arrived from Sigma, and I have created stock and working dilutions.
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