Thursday July 15, 2010

Raida

• Today I did a Colony PCR of the the I0050 + B0034 Construct with the primers that Emily provided me with.

PCR conditions: Stage 1: 94 degrees Celcius Stage 2; Step 1: 94 degrees 1 minute Stage 2; Step 2: 55 degrees 45 seconds Stage 2; Step 3: 72 degrees 1 minute 15 seconds Stage 3: 72 degrees Holding Temperature: 4 degrees Celcius

• I have also assisted Alex with his Gel electrophoresis of the CpxP colony pcr. We are hoping to see a band around the length of 150 bp.

Jeremy's restriction enzyme on pRFP EcoRI/HindIII and XbaI/HindIII to remove pRFP from its original plasmid

Jeremy

• Today I reconstructed K239000 with I13507 and I13504, then K135000 with I13507 and I13504 with proper plating. I also did a restriction enzyme digest of pRFP with EcoRI and HindIII and also XbaI and HindIII. And a gel was run of the restriction enzyme products. Also 4 tubes were prepared for I0500+B0034+E0040 to test inducing arabinose.

Alex

Ran all three gels of the PCR reactions. The reaction containing the highest magnesium ion concentration yielded PCR in all 12 reactions. The DNA found was of the right size (~200)bp.

Himika

• Today I did more research about high and low copy plasmid. Dave came up with a new idea of constructing the transcription/translation circuit and Emily and I spoke about it to finalize a strategy.

Chris

Today, I did more calling of companies to apply for sponsorships as well as writing the application for Transalta sponsorship. We also did PCR purifications of the CpxP promoters but ended up with very low concentrations and very high levels of contamination with proteins and RNA. Because of this, we decided to redo the PCR purifications but ended up with little difference in concentration and contaminations. Regardless, we did restriction digests and left these overnight to place into plasmids tomorrow.