Lab Notebook

14 August 2009

Sriram

I found that colonies grown from plates were very less (around 1 to 5). There must be something wrong with the assembly. I must run the gel on monday to check the digesion. Also i found that pLacI, pTet and pSB1C3 DNA were getting less hence I must prepare them also on monday.

Tim Weenink

First did ligation of !E and !F with the parts that were not excised from gel.

Then I did electroporation of the !E and !F gel extracted assemblies.

After that I did chemical transformation of !E and !F assemblies (both gel extracted and direct PCR restricted fragments)

Calin

oriTR_KO +L-ara +PCR and trbK_KO +L-ara +PCR backup plates made on 0.5x KAN.

Did miniprep on CB and CC culture tubes.

Plate 2, 4, 6, and 8 are overgrown. Cells survived electroporation. No colonies on knockout plates or control plates.

Ran 1.5% gel with CC and CB.

Digested CB with X + P (11uL DNA). Digested CC with E + S (4.5uL DNA).

Daniel

No growth although gel seems good. Let´s think about it during the weekend