NextSeq 500 sequencing; Parallel monitoring of the integration efficiency and stability of mini-TLCΦ plasmids with different attachment sites
Parallel monitoring of the integration efficiency and stability of mini-TLCΦ plasmids with different attachment sites
FX492_s
β2163 cells harbouring the mini-TLC plasmids were collecting  on LB plates supplemented with Dap (0.3mM) and chloramphenicol.  genomic DNA extraction kit Plasmid attP sites were amplified by performing 17 PCR cycles on 50-100 ng of plasmid library gDNA with an equimolar mix of 4103-4105 P5 and 4178-4180/4246-4248 P7 primers. The products were purified from the P5 and P7 primers the double selection with AMPure.