Lab Notebook

17 August 2009

Calin

The backup trbK_KO pate has 11 colonies. The Plate 3L oriTR_KO -L-ara +PCR also had 8 colonies. No growth on any other plates. Plates with growth put in fridge.

Ran colony PCR on 5 Colonies: Did colony PCR for 2 different trbK_KO colonies and also made 5mL culture tubes for them. Also did colony PCR on the oriTR control and 1 R751+pKD46+trbK_mod_S and 1 R751+pKD46+oriTR_mod_S for reference.

Made 100uM and 40uM stock of verification primers.

Made 1 L LB agar.

Made 10mL of TRIMETHOPRIM stock.

Tim Vos helped me transform CA into R751 Electrocompetent cells. Both plate and 5mL cultures made.

Ran a 1% and a 2% gel with all 5 colony PCR samples.

Daniel made 10 TRI, 4 CAM, and 6 AMP plates.

I made a further 15 AMP + CAM plates.

Tim Weenink

checked number of colonies in transformation plates: !EEgelex: 1 colony !FEgelex: 0 colony !ERnonex: 1 colony !FRnonex: 2 colonies (+one on lid?) 1ERgelex: 0 colony !FRgelex: 0 colony

did colony PCR on all obtained colonies and the weird opaque drop on the lid.

ran gel of the PCR products: see tomorrow for image.

Inoculated *2S and !SA cultures and induced them with IPTG for protein gel. (also some without induction for control) Saeed helped with this.