Newcastle University Wetlab/14 August 2008

Thursday 14th August

 * BBa_I746101 and BBa_746107 were isolated and run on a gel together with the 4 restrictions performed 13.08.08.

Lane 1: 1kb ladder

Lane 2: empty

Lane 3: pGFPrrnB restricted with EcoRI + NheI

Lane 4: pJWV021 restricted with NheI + BglII

Lane 5: pUC57-ncl08 restricted with EcoRI + NheI

Lane 6: pUC57-ncl08 restricted with NheI + BglII

Lane 7: BBa_746101

Lane 8: BBa_746107

The gel showed that there was very little pGFPrrnB (restricted with EcoRI + NheI) and no pJWV021. It was concluded that the 2 step restriction and purification method reduced the plasmid yield significantly. As it was later discovered that this method was unnecessary (as NheI and BglII are compatible using buffer M), it was decided to perform the restriction as a single step in future.

The gel also showed successful restrictions of the pUC57-ncl08 plasmid and lots of the two BioBrick samples.


 * The 2.2kb fragments from the pUC57-ncl08 restrictions were cut out of the gel using a UV light box and scalpel blade (see Cutting a Specific Band from Agarose Gel). The resulting gel slices were then purified to remove all gel and enzymes (see Purifying DNA from Gel Slices).