Team:Groningen/Template/MODULE/Notebook/toolbox/week10

Because of the constant low concentrations of the plasmid pSB1C3 with the new BioBricks, it was decided to transform the isolated pSB1C3 with the new BioBricks (NisA, NisC, NisRK, PNisI and sfGFP(Bs)) again in Escherichia coli DH5α. The pSB1C3 plasmids containing the new BioBricks were again isolated. For each transformation, two seperate cultures were grown in LB with 10 μg/ml chloramphenicol. A miniprep was done on the cultures and the insert size was checked with colony PCR, using VF2 and VR as primers. In addition, a restriction analysis was done using EcoRI and PstI. Cultures that showed a correct insert size were sent for sequencing.