EpsE, 5' and 3' AmyE integration sequences digested with EcoRI and PstI
U = undigested, D = digested
All the inserts for 3' AmyE and EpsE (above) and 5' AmyE (below) appear correct.
EpsE Single Colony PCR reactions were cut with HindIII to check that the insert was EpsE and not the possible contaminant, RFP-Terminator
U = undigested, D = digested. EpsE on the left is cut with HindIII, 5' AmyE cut with EcoRI and PstI
The insert in minipreps one and two (numbered left to right) is has lost a small fragment (~150bp) when cut, as should be the case with EpsE which possess a HindIII site about 100-150 away from the 3' end of the gene. The third miniprep shows what appears to be two fragments (one approximately 250bp, the other approximately 400bp), though it is more likely that there are three, as would be the case if RFP-Terminator was the insert instead.