Brown: Team Resistance/22 October 2008

22 October 2008

Ligation

pVJ4 lysis cassette and S105 lysis cassette into standard biobrick vector plasmid pSB1AK3 at EcoRI and SpeI

• 15 ul mixes:
• 12.37ul diH20
• 1.5ul T4 10x buffer
• 2.08ul pVJ4 OR

2.62ul S105

• 0.53uL vector
• 0.5ul T2 ligase

Transformation

• Used 50ul competent cells (invitrogen)
• skipped the SOC step (immediately plated) since SOC step not necessary for amp resistant plasmid
• Put in incubator at 10:30am

Conductance test

Measuring conductance change and optical density change in parallel

Set up two cultures of the pVJ4 plasmid

1. control (no ara*)
2. test (480ul ara*)

Because the cells grew during the experiment into stationary phase, the lysis seems to take much longer than usual.