21st July

Sriram

Finally transformation worked in 5 plates out of 6 and for unknown reasons the pBla biobrick was not transformed and not seen in the plate. At 5:45 PM the best colony from 5 plates were cultured in 5 ml culture tubes with respective antibiotics and incubated at 37 °C and 160 rpm. So tomorrow we can proceed to the DNA extraction step.

Found that the competent cells must be prepared. Tim Weenink has already cultured top 10 cells for competent cell preparation. I diluted the culture in 100ml flask and grown till 0.49 OD at 600nm. While the culture was growing I prepared the TSS buffer and filter sterilized with 0.2micron syringe filter and stored in freezer. Then 50ml culture was split into two falcon tubes and centrifuged. After throwing the supernatant it was resuspended each tube with 2.5ml TSS buffer.

Calin

Got transformed colonies in all 4 plates. I cultured the best colony in each plate in were cultured in 5ml culture tubes with 1x ampicillin added and incubated at 37 °C and 160 rpm at 5:30PM. So tomorrow we can proceed to the DNA extraction step.

Tim Vos

Since Sriram became busy with culturing the transformed colonies, I made 40x100 µl aliquots into cool and sterile eppendorfs in laminar flow.