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Team:British Columbia/Notebook
From 2013.igem.org
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Team CRISPR
July 3
List of BioBricks that are/will be on the way:
Cas9:
- Cas9 - Joe and Anna
- pTet -
- const. promoter + Cas9 - Joe
- promoter + Cas9 + Leader + R + spacer + R (on Kan)
- L + R + spacer + R
Target:
- tracerRNA
- repeat + spacer + repeat - Vanins*2 + underlings
- leader - Ray, Fisal, Dan
Decoy:
- PAM + spacer (on Amp)
Team Cinnamaldehyde
4-coumarate-CoA ligase:
- PCR worked
Team Caffeine
June 20
Joe, Dan, and I made heat-shock competent cells today - they can be found in the top compartment of the -80º Freezer #3 in the room full of freezers (thanks Cam!). We will be ordering primers (through Diane) to modify the TU Munich caffeine biosynthesis BioBricks we got in the Distribution Kit.
We're making a start - what are the other groups up to?
- Grace Yi
June 27, 2013
TU Munich Parts that we received were transformed into these competent cells, and transformation efficiency was very low, giving us two colonies per plate. The three parts (17H, 17J, 17L) were miniprepped, with DNA concentration of 154.3ng/ul, 323.8ng/ul and 117.3ng/ul respectively.
- Liz Geum
July 4, 2013
We ran an initial PCR on the miniprepped plasmids with biobrick primers (VF2 and VR) to confirm the presence of the parts. Gel confirmation of the PCR products showed a band near 1kb, our expected gene size, for one of the genes only. The other samples did not show any bands. Because the transformation efficiency was very low, we decided to troubleshoot transformation with new competent cells we received from Ray, our grad advisor. We also designed with Ray and ordered the"stitching" primers that would allow us to assemble our biobricks easily without having to standard-assemble. In the design, we included a consensus sequence to allow for polycistronic transcription.
- Liz Geum
July 9, 2013
Transformation with new competent cells was highly efficient.
