Team:Groningen/Labwork/6 September 2013



The plates (pSB1C3-S1-S5 and pSB1C3-S2-S5) show single colonies.
ColonyPCR was performed on 6 colonies per plate (total of 12 colonies) beside one positive control (pSB1C3-S1).
The primers used were VF2 and VR (annealing temperature 56°C).
The samples were checked on agarose gel 0.8%.

pSB1C3-S1-S5 colonies.

pSB1C3-S2-S5 colonies.

Two colonies seem to be positive candidates.
Colony A from the plate pSB1C3-S1-S5 was inoculated over-night in liquid culture (LB + Cm).
Colony C from the plate pSB1C3-S2-S5 was inoculated over-night in liquid culture (LB + Cm).


All inoculations form yesterday grew.
After spinning down for plasmid isolation, some pellets were green/red (indicating that the promoter is inserted right)

Row1: GFP0804 (1-4) pXs1s13 (5-8)
Row2: P:RFP (1-4) GFPdsm (5-8)

Did an restriction check on pXs1s13 (silk) with EcoRI, analysis of the gel showed that the silk did not ligate into the pX plasmid.