Newcastle University Wetlab/28 August 2008

Thursday 28th August

 * New frozen glycerol stock of cells made to be kept at -80˚C.


 * Isolated plasmid from overnight culture of pGFP-rrnB. For this the 10mL culture was divided into 5 x 2mL plastic tubes and pelleted by centrifugation for 10 minutes. The supernatent was discarded and follwing the isolation procedure the product was pooled.


 * Restrictions of pUC57-ncl08 and pJWV021 were carried out.



Lane 1: 1kb ladder

Lane 2: pUC57-ncl08 restricted with EcoRI

Lane 3: pUC57-ncl08 restricted with BglII

Lane 4: pUC57-ncl08 restricted with EcoRI and NHeI

Lane 5: pUC57-ncl08 restricted with NHeI and BglII

Lane 6: pUC57-ncl08 control (no enzyme)

Lane 7: pJWV021 restricted with NHeI and BglII

Lane 8: pure pGFPrrnB


 * This gel shows that only partial restrictions of the plasmids had occurred. It was subsequently discovered that an incorrect quantity of buffer was being used. This was rectified in further restriction reactions.