Team:Groningen/Labwork/29 July 2013

Mirjam

Run the samples of the colony PCR of BBa_K823823 combined with the LacI promoter made on 27-07 on gel (order 1-8).
There are no bands around 1500 bp.

Run the purified PCR product of spec on gel.
The purified spec shows a nice band.

Run the PCR products of silk 2 on gel to do gel purification.
Only one of the tubes showed a band around the expected height.

Run the PCR products of silk 2 on gel to do gel purification.

Made a double restriction digestion with fast restriction digest for Pdes and CheY with respectively EcoRI and BamHI; and BamHI and PstI.

Chaline

Made a ligation for the knockout system of Des, with Des UP, Tet and Des Down

Friso

Made a restriction digestion for Spec purified.

Claudio

Two GFP reporters are picked from the iGEM database: BBa_E0840 and BBa_E0240 (backbone: pSB1C3). Those are transformed into E. Coli DH5Α (Cm resistance).

The plasmid containing mKate2 (red fluorescent protein into pGEM-T Easy vector) is inoculated in LB + Ampicillin resistance.

The Colony PCR which failed on Saturday is performed again on 4 colonies (the same which were also tested the previous time) changing the annealing temperature to 46°C.
The four samples are loaded on gel and bands are spotted around the expected height (~1500 bps).
The transformation worked.