Lab Notebook

17 August 2009

I cultured pLacI, pTet and pSB1C3 biobricks for miniprep tomorrow. I ran gel of the previous restricted parts for the assemblies I did on thursday (13/08/09). The image we checked needed more staining hence we kept it for overnight staining. The gel image is in next page.

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.

Daniel

During the weekend I noticed I need more biobricks. Besides, I wrote emails to people involved in previous iGEM´s teams of Berkeley, Clathec and Peking which had worked with riboregulator regarding to the synthesis of locks and keys.

Rehydratation and transformation of C0051, P0440, R0051 and I13504. After that culture in plates and tubes.