Uppsala-Sweden/15 September 2009

From 2009.igem.org

Picking colonies and colony PCR

The colonies from the plated transformations since yesterday were picked this morning, together with the anti sense RNA's that didn't seemed to have worked according to the evaluation gel from yesterday.

pPetE-E1A1-T, pPetE-E1B1-T, pPetE-E1B3-T, pPetE-E21-T, kivd, RBS-pdc-RBS-adh2z, RBS-kivd, kivd-RBS-YADH2-T, pPetE-RBS and RBS-PirA-RBS-PirB-T.

These colonies were run through a Taq PCR.

RK = Rbs-Kivd, RPRA = Rbs-Pdc-Rbs-Adh2Z, RART = Rbs-PirA-Rbs-PirB, KRYT = Kivd-Rbs-YAdh2-Ter

Some strange extra bands, probably due to annealing of the verification primers to the RBS as they share a high sequnce simalarity.

As seen by the gels we have working colonies of pPetE-Rbs, Kivd, P-E1A1-T, P-E1B1-T, P-E1B3-T, P-E21-T, and RPRA. These were inoculated for plasmid prep tomorrow.

--Flormane 12:54, 15 September 2009 (UTC)

Glycerol Stocks

Glycerol stocks were created for teh aRNA and the RPRAT construct.

Plasmid Prep

Plamidpreps were performed on previously [2009-09-14] inoculated aRNA constructs. Concentrations 100-150 µg/ml.

RPRAT [uncertain version] plasmid prepped.

Transformation pPetE- R0011- pTRC- -RPRAT to pSB1AC3

Utilizing our standard protocol the promoters pPetE, R0011 and, pTRC were ligated together with RPRAT into the AC3 plasmid.

pTRC seems empty, pPet E badly digested (single and double)RPRAT looks alright. --Anders 20:20, 19 October 2009 (UTC)

Gel Colony PCR of GFP and Lux System Parts

Ai: A340620 + i13504 (2,1kb) AJ: A340620 + J37034 (2,8kb) ACO: A340620 + C0261 (1,9kb) AB: A340620 + B0034 (1,2kb) Pi: pPetE + i13504 (1,6kb)