Team:Newcastle/IntroductoryLabwork/6 July 2009

Introductory Lab Session: 6th July 2009

Goksel rehydrating the BioBricks, which are in dry DNA form

List of bricks for the day

We got 3 tubes containing cells from the freezer at 80C and placed them into the ice and waited for 30 minutes
We prepared 6 tubes (3 tubes for controls and 3 tubes for the plasmids) for the bricks
We added 40ul of cells to each of the six tubes
We added 3ul of DNA for the plasmid tubes and 3ul of water for the control tubes
We added 900ul of LB for each of the six tubes
We then placed the tubes in the shaking incubator for an hour.

James extracting portion of diluted solution to be placed on plates

Hanny spreading solutions on plate using sterile glass beads

We prepared 9 plates with full strength (undiluted) and 9 plates with diluted tube content.

The first three plates having LB + Amp were used as the negative control. Hence the cells not having amp. resistance would not grow on these plates.
The next three plates with LB content were used as the positive control. Cells without amp. resistanec would still grow on these plates.
The other three plates were used for the plasmids.

LB + Amp, (-) control 1	LB, (+) control 1	LB + Amp, Transformation Plasmid 1
LB + Amp, (-) control 2	LB, (+) control 2	LB + Amp, Transformation Plasmid 2
LB + Amp, (-) control 3	LB, (+) control 3	LB + Amp, Transformation Plasmid 3

For the diluted ones we used 900ul of LB and 100ul of DNA from the corresponding tubes.
The same set of 9 plates were prepared for the diluted content.