TUDelft/11 September 2008

=September 11th=

Transformation of PCR products
Ligation of PCR products of E. coli genes was performed o/n and a direct transformation of the ligation products was performed. 50 ul of competent cells were added to the 15 ul ligation mix transformed o/n. Results will be checked by colony PCR tomorrow.

Colony PCR of 3A Assembly
The transformants from the assembly have grown, indicating the assembly has worked. We did a colony PCR on a couple of the transformants, to check for insert sizes. The gel of the colony PCR is displayed in figure 1. We suspect the parts which gave multiple bands at very different sizes are parts which have not ligated inserts but are just a couple of backbones ligated together, as this would give primer annealing zones in all kinds of orientations. For each assembly we've PCR'd a construct of the correct size (some extra bands could be a consequence of the use of B0015 as terminator).