PCR testing for insertion

We want to test for gene insertions and recombinations at several steps of our project. PCR is the easiest and fastest way to determine the success of Recombineering.

galK insertion

Using a set of primers flanking the galK cassette, we tested the insertion on our recombinants. galK is around 1200 base pairs in size, so we would expect the recombinant PCR to be about 1200 bp larger than the non-recombinant. Indeed, this is exactly what we saw.

SDS-PAGE Protein Array

Hcp secretion is the hallmark of a working T6SS. We plan to use SDS-PAGE to confirm our promoters.

Competition Arrays

Once we have a working T6SS in E. Coli, we can use GFP-labelled organisms to test the competitive advantage of our T6SS strain vs. non-T6SS organisms.