Lab Notebook

30 July 2009

Sriram

Prepared the chemically comepetent using both TSS Buffer protocol and TMF Buffer protocol. Altogether it must come for 200 transformations if all works. Along with Calin I prepared the electrocompetent cells which was a bit laborious but we managed to prepare them.

Tim Weenink

prepared assembly mixes for !B, !C and !D constructs. Standard Ginko assembly kit was used. DNA concentrations as below (these mixtures were added to 7.5 µl of digestion mixture):

Ligated and transformed assemblies according to protocol.

15:20 Done plating the assemblies.

15:40 inoculated *S and *T cultures, four colonies each

16:55 Inoculated !A cultures from glycerole stock. Two tubes.

Calin

Plated three backbones left over from the transformation the other day. Did a digest on oriT-R with E and P. Made 3 x 5ml culture tubes for the pSB4C5 backbone which had a few colonies. Made electrocompetent cells with the [http://openwetware.org/wiki/Knight:Preparing_electrocompetent_cells Knight:Preparing electrocompetent cells] protocol. Flash froze tube in liquid nitrogen. Checked backbone concentrations:

Ran gel on backbones and oriT digest. Gel left for too long.

Tim Vos

Minipreped 4 x pSB1C3, 4 x pSB4A5, 2 x rbs-GFP, and 4 x pSB1A3. One pSB4A5 tube is incorrect, it shows RFP expression. Also loaded the gel.

Orr

Made some chemically competent cells using the TSS buffer protocol. Started making a Java program that will allow lock sequence design based on the chosen RBS, and will design the equivalent key based on the lock sequence. The program will incorporate a database with all the working RBS segments in the iGEM registry, so that the user can choose which RBS he wants to use.