Joyce
Plasmid prep of the overnight cultures
ND1000-nanodrop measurements: DNA concentrations are between 30 and 60 ng/μl
Inoculating 2 erlenmeyers of 200ML , 1 for making competentcells DH5alpha and other one for TOP10 cells
Also TOP10cells will be send to the iGEM Wageningen team.
Also preheat other 2 200ml erlenmeyers
Restriction analysis:
Digestion test:
2μl FD buffer
0.5μl EcoRI
0.5μl SpeI
8μl MQ water
10μl + 10μl sample
Digestion mix:
30μl FD buffer
7.5μl EcoRI
7.5μl PstI
120μl MW water
Digestion mix:
30μl FD buffer
7.5μl XbaI
7.5μl SpeI
120μl MW water
Work in Clone manager: check primers PhybB again
Jakub
Week 24:
- cloning double terminator into chloamphenicol, ampicilin, kanamycin and tetracyclin backbones
Christoph
- Repeated cloning of reversed PLasB into pSB1A3+DT.
- Cloned PcI into pSB1K3+rev.Terminator