Team:UNIPV-Pavia/Calendar/June/settimana5

June, 28th

Planning of the activity of the week. Freezer cleaning: for each ligation, we chose the correct clone and stored it in the iGEM 2010 ligations box. All these cloned were gel screened and sequenced and are correct! The colonies we chose are:

Other colonies resulted positives at the screening but were NOT sequenced and are stored in the iGEM 2010 cemetery box. These clones are: I1-1, I2-2, I4-1 and I6-3.

June, 29th

Inoculum of I6, BBa_J23118, BBa_J23110, BBa_J23114, BBa_J23116 from glycerol stock in 5ml LB+Amp. Cultures were grown ON at 37°C 220rpm.

June, 30th

PhaP-1 and PhaP-2 plates showed colonies!! A colony from each plate was picked and used to infect 1ml LB+Kan. Liquid cultures of PhaP-1 and PhaP-2 were grown for 6 hours at 37°C 220 rpm and then used to prepare glycerol stocks. Remaining liquid cultures were re-filled with 5 ml LB+Kan an grown ON at 37°C 220 rpm for tomorrow MiniPrep. Glycerol stocks of PhaP-1 and PhaP-2 are stored at -80°C in the iGEM 2010 Registry box.

Ligations were heated at 65°C for 5 minutes to inactivate ligase ad then tranformed in E. coli TOP10 home made competent cells. Plates were incubated ON at 37°C (we decided to incubate plates 5 hours longer than usual, because cell growth was already slower for I6).