From 2006.igem.org

Thursday 7 September 2006

We grew up M30109 last night and I removed it from the inc at 11:25 am this morning. Next, we need to culture it in LB Amp+Clm broth, miniprep it, and then electroporate it into EnvZ- and WT strains of E. coli.

Culturing

1. we need to make LB stock broth (2 500 ml bottles)
2. we need Ampicillin stock
3. we need Chloramphenicol stock

Then we can make 5ml of LB Amp+Clm broth in a 5ml tube to culture our E.coli containing M30109.

protocols

See this [http://openwetware.org/wiki/Bacterial_cell_culture OWW protocol on making cultures] for reference when making the broth.

I'll post a little protocol for making antibiotic stocks later.

Miniprep

protocols

Electroporation

protocols

We used [http://openwetware.org/wiki/Knight:Electroporation Tom Knight's electroporation protocol] last time and it worked great.