Team:Groningen/Labwork/16 July 2013

From 2013.igem.org

Mirjam
Finally the colonies are growing on LB agar plates with ampicillin. The negative control does not show any colonies, the double negative does??, the positive control shows a lot of red colonies. On one of the four plates there are no colonies present, on two of them only a couple and on one a lot.

The colonies are picked and placed in LB medium with ampicillin to grow further for miniprep.

Inne
Did a PCR with 3 samples in duplo
sample 1: primer F without strep tag
primer R without strep tag
annealing temp: 78 C
sample 2: primer F with strep tag
primer R without strep tag
annealing temp 78 C
sample 3: primer F without strep tag
primer R with strep tag
annealing temp 80 C

Protocol:
28.2 uL MillQ 1.5 uL 5% DMSO 1 uL of each dNTP 10 uL GC buffer 2.5 uL primer F 2.5 uL primer R 1 uL template DNA bba_16 (Utah silk biobrick) 0.3 uL phusion polymerase

Sander
did a Restriction digestion for Silk and Signal Sequences. incubation started at 11:00 for the signal sequences and 11:30 for the silk.