Analysis of the cloning of OmpR and EnvZ

PCR screening

• positive control: S158 (pSB3K3)
• negative control: no template
• screening PCR programm
• elongation time: 1 min 30
• primers: O18 & O19

Reaction mixture (25 µL total volume)

• 12,5 µL Quick load PCR Mix
• 0,5 µL O18
• 0,5 µL O19
• 12 µL water

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.

Miniprep and stock glycerol of L151

• Protocol stocks

Stock number	Ligation name	Biobricks	Description
S162.1	L151.1		rbs-LasR - Double terminator
S162.2	L151.2
S162.3	L151.3

• Protocol minprep

Miniprep Name	Ligation name	Antibio	Biobricks	Description
MP162.1	L151.1	Kan		rbs-LasR - Double terminator
MP162.2	L151.2
MP162.3	L151.3

Digestion of MP162

Measurement of the concentration of MPs 162

Digestion

protocol