Today minipreps were prepared. These were digested and run on a gel to confirm the presence of the correct constructs. The folowing constructs were minipreped: Gene art 14 (pXyl-spoVG) and 5' EpsE integration sites.
To test that insert with the correct size is present in the plasmid, the minipreps were digestd with EcoRI and PstI. The image below shows the results:
Several of construct 14 appear to have succssfully been ligated. In addition all of the 5'EpsE appear to have worked.
To construct 5'EpsE integration site we used a PCR product and as a result of the approach, there is a possibility that the orientation of this biobrick in the vector is incorrect. Digestion with XbaI SpeI will confirm correct orientation.
If the PCR product has the correct orientation then the insert will fall out with this digestion. Clearly only 5'EpsE-1 is correct.