Newcastle University Wetlab/4 September 2008

Thursday 4th September

 * The plates from 03.09.08 showed expected colony growth and therefore overnight cultures were made from individual colonies on Plates 1, 2 and 9. The culture media were made using 10mL of LB in 15mL plastic tubes together with the correct volume of the relavent antibiotic. This was 5μl spectinomycin for the GFP-rrnB colonies and 4μl kanomycin for the pJWV021 colonies. Cultures were taken as follows:

GFP-rrnB - pUC57 ligation (grown on spec)
 * 9 white colonies (1-9) (Plate 1)
 * 1 green colony (10) (Plate 1)
 * 2 white colonies (11,12) (Plate 2)

pJWV021 - pUC57 ligation (grown on kan)
 * 10 white colonies (13-22) (Plate 9)
 * 1 pink colony (23) (Plate 9)
 * 1 bright white colony (24) (Plate 9) (this is likely to a be contaminant non-E.coli colony)


 * All 24 culture tubes were incubated at 37˚C whilst shaking for ~20 hours. This large quantity of cultures should ensure that we obtain at least 2 cultures (one GFP-rrnB and one pJWV021) that have taken up the ncl08 insert. The plates were kept at room temperature in case fulrther stab cultures needed to be made.