Team:Groningen/Labwork/14 September 2013

Claudio

Plates containing BBa_K1085014-S1-S5-S14 and BBa_K1085014-S2-S5-S13 showed around 20 colonies.
Colony PCR (NEB Master Mix) was performed on 4 colonies per plate using F_spank_NheI and HM14 primers (annealing temperature 65°C).
The samples were checked on agarose gel 0.8%.

Colony B from plate BBa_K1085014-S2-S5-S13 showed a band at the expected height (~2400 bps). Therefore was inoculated in LB + Cm.

Plates containing pSB1C3-S1-S5-S13 and pSB1C3-S2-S5-S14 showed thousands of colonies.
Colony PCR (NEB Master Mix) was performed on 4 colonies per plate using VF2 and VR primers (annealing temperature 65°C).
The samples were checked on agarose gel 0.8%.

Colony C from pSB1C3-S1-S5-S13 and colony A from pSB1C3-S2-S5-S14 show to be positive candidates and were inoculated in LB + Cm.

S13 and S14 were ligated into pSB1C3-S1-S5-S5 and pSB1C3-S2-S5-S5, respectively.
S13 and S14 were ligated into pSB1C3-S16-S3-S5 and pSB1C3-S16-S9-S11, respectively.
The ligation products were plated on LB + Cm.

Sebas

Checked if the streaked plates from yesterday grew. And restreaked the positive colonies on a amp/iptg/agar/LB plate.