Team:Paris Liliane Bettencourt/Notebook/2011/09/01/

From 2011.igem.org

Cyrille

Troubleshoting K606003

The glycerol was stricked on a plate. The region where the plasmids are too concentrated loose the plasmid.

Miniprep of pHM3 from glycerol

Two miniprep of pHM3 where done.

Cloning of YFP-TetR

The strain K1 (containing LacO) was open in SP. The digestion was PCR Purified.

The pHM3 was digested in EP and gel purified, but it didn't gave any yield.

There where very few of YFP-TetR left. The digested product was not visible on the gel.

Launch again YFP-TETR culture

The glycerol YFP TetR was launched again in 3 tube for miniprep tomorow

Comptetent cell debbuging and preparation

There where ampicylin contamination in comptetent cells. Turbocells where stricken on a plate for competent cell preparation tomorow.

Danyel

GFP amber

Quick change mutation was launched last night of: GFPmut3b (E0040) and GFP + ter with the same primers. Today, PCR products were digested with DpnI FastDigest enzymes and run on gel.

Ladder_Blank_GFP+ter: control_20ng_40ng_blank_GFPmut3b control_20ng

tRNA amber suppressor & T7 amber

S81: Pveg+spoVG+tRNA+ter S80: T7amber + ter S24: Pveg+spoVG S38: tRNA in pSB1C3 S58: S38 + ter

Yesturday, S81 was ligated into pHM3 and S80 was ligated into S24. Both constructs were transformed and plated, then left to grow overnight. The negative control of pHM3 transformation only gave one colony for 20µL insert:vector = 6:1 The negative control of S24 gave innumberable colonies. Nonetheless, a PCR colony was launched for both.