IGEM:Groningen/Notebook/iGEM 2011/2011/08/15

From OpenWetWare

Jump to: navigation, search
Project name Main project page
Previous entry      Next entry

15-8-11




colony PCR of taRNA-pSB1A3-DT:
Composition master mix:
10× Taq buffer: 40μl
dNTPs 10mM: 8μl
MgCl2: 24μl
Forward biobrickvector primer: 8μl
Reverse biobrickvector primer: 8μl
Taq polymerase: 2μl
MQ water: 310μl

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 30s
Final extension: 72°C for 10min
Store infinite at 4°C
Analyse PCR samples on a 1% agarosegel

Check sequencing results PBAD-RBS-GFP-DT
again the GFP part does not look well
Filling in 'stageformulieren'
Grow TOP10 competent cells in M9 minimal medium and also E.coliDH5alpha, to see tomorrow if the TOP10cells grow better
compared to the E.coli DH5alpha cells...
Cleaning up the lab for tomorrow's audit!


Personal tools