Team:Paris/August 18

=Analysis of the cloning of OmpR* and EnvZ*=

PCR screening
Reaction mixture (25 µL total volume)
 * positive control: S158 (pSB3K3)
 * negative control: no template
 * screening PCR programm
 * elongation time: 1 min 30
 * primers: O18 & O19
 * 12,5 µL Quick load PCR Mix
 * 0,5 µL O18
 * 0,5 µL O19
 * 12 µL water

Minipreps & Stocks
We wanted to analyse the transformants of the ligations L133 (OmpR* in pSB1A2) and L134 (EnvZ* in pSB1A2) from the experience of august 9th/11th. 3 clones of L133 and 3 clones of L134 were analysed. Each clone tested was then cultured in 7,5 mL LB + ampicilline.

=Construction of rbs-LasR-dble ter=

Miniprep and stock glycerol of L151 (rbs-LasR - Double terminator)

 * Protocol stocks
 * Protocol minprep

=Construction of : promotor-rbs-LasR - Dble ter=

Digestion
Protocol

Gel Extraction
Protocol

Ligation
Protocol

=Culture of stable strains with biobricks 2008= Protocol