15 October 2008

Wet Lab

Digestion of minipreps

• 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.