IGEM:Groningen/Notebook/iGEM 2011/2011/08/26

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26-8-11



colony PCR of PhybB-RBS-cI-LVA-DT, PhybB-RBS-LasR-LVA-DT and PBAD-cI-LVA (3 colonies):
Composition master mix:
10× Taq buffer: 48μl
dNTPs 10mM: 9.6μl
MgCl2: 28.8μl
Forward biobrickvector primer: 9.6μl
Reverse biobrickvector primer: 9.6μl
Taq polymerase: 2.4μl
MQ water: 372μl

PCR conditions:
Pre heated lid: 111°C
Denaturation: 94°C for 10min
Cycle (33×)
Denaturation: 94°C for 30s
Annealing: 60°C for 30s
Extenstion: 72°C for 2min
Final extension: 72°C for 10min
Store infinite at 4°C
Analyse PCR samples on a 1% agarosegel
7 out of 9 for PhybB-RBS-cI-LVA-DT have the correct size and 5 out of 9 for PhybB-RBS-LasR-LVA-DT have the correct size!
Send them monday for sequencing

Plasmid prep of PBAD-RBS-GFP again.
ND-1000 Nanodrop measurements: plot looked better than before.
50 ng/microliter DNA was obtained.
Samples were prepared and send out for sequencing.

Teammeeting at 14:00!