Team:TUDelft/20 July 2009

=20th July=

Sriram & Calin
No colonies on the plates from Friday. Suspect either that the 300 rpm mixing during the heat shock might have killed the cells, or that we added the cells too soon after adding the antibiotic, before it had time to diffuse. Redid the transformations correcting these steps once again with the following parts: For Conjugation: BBa_I714031 - OriT-R BBa_J23100 - strong promoter BBa_E0840 - GFP generator BBa_I13522 - pTet GFP For Delay: BBa_E0040 - GFP BBa_C0040 - TetR BBa_C0051 - cl BBa_I14018 - pBla BBa_I12006 - pLambclin BBa_E1010 - mRFP1

Heat shock was done in the waterbath at 42&deg;C for 1 min. Antibiotic was added while the agar was still liquid, mixed, then poured onto the plates to solidify.

A beaker of eppendorf have been sent to kitchen for autoclaving.

Weenink
I did the assembly of my first biobrick today! Protocol was according to the ginkgo bioworks assembly kit. The following biobricks were used: Upstream: B0032 Downstream: K145015 Backbone: pSB1AK3 Transformation was done on the PCR machine (as a test) Also, I inoculated 5 ml liquid culture (lb, 37degrees shaking incubator) of top10 cells for making them chemically competent.