Team:Groningen/Labwork/5 September 2013
From 2013.igem.org
Claudio
The plates (pSB1C3-S1-S5 and pSB1C3-S2-S5) show a lot of colonies.ColonyPCR is performed on 4 colonies per plate (total of 8 colonies) beside one positive control (pSB1C3-S1). The primers used are VF2 and VR (annealing temperature 58°C).
![](https://static.igem.org/mediawiki/igem.org/0/06/ColonyPCR_pSB1C3-S1-5_pSB1C3-S2-5.jpg)
Colony B from pSB1C3-S1-S5 and colony D from pSB1C3-S2-S5 seem positive candidates.
Those two colonies were streaked out on LB + Cm and incubated overnight at 37°C.
PCR was performed on S3 and S9 using primer pair S34910.
PCR was performed on S6, S13 and S14 using primer pair S16 (Sander).
PCR was performed on S16 using primer pair true-S16.
The annealing temperature tested for all of them was 58°C.
The samples were checked on agarose gel 0.8% (Sander).
The PCR worked for all the templates.
The PCR reaction were purified (Sander).
Sebas
All plates had colonies.Restreaked 8 colonies and inoculated 4/8 colonies in 3ml LB/amp.
Performed a colony PCR on pX+s1s13 (colony 1-4).
(lost sample due to gel problem)