Team:TUDelft/14 July 2009

=14th July=

Lab work: Sriram & Orr
Prepared 1 litre of LB agar medium and sent to Kluyver lab's kitchen.

Using 650 &micro;l of competent cells (E.coli DH5alpha) and the transformation protocol, we inserted 13 biobricks [R0010, I14018, I12006, R0040, J23008, J23031, E0040, E1010, C0051, B0034, B0015, S03335, S03473, K081013 and C0040] for delay into competent cells and grew them in a solid media to increase the amount of biobricks we will have.

Diluted the 13 biobricks with 15 &micro;l RNase free water and stored in -20°C freezer.

Standard transformation procedure:

Remove competent cells from -80&deg;, let thaw for 10 min on ice and aliquot in 50 &micro;l amounts. add 2-5 &micro;l of vector, to 50 &micro;l cells, no mixing by pipet due to shear induction. keep on ice for 20 minutes (vector spreading through volume). heat shock (42°C) for 45 seconds. keep on ice for 2 minutes. add 200 &micro;l SOC, put on 37°C for 1 hour or longer with agitation (160 rpm). plate out 250 &micro;l on appropriate antibiotics.

Weenink
Brought beaker of eppendorf tubes to autoclaving

Calin
Looked into the details of lambda red knockout protocol. Started designing the required primers.