Wisconsin: Lignin Project/7 July 2008

From 2008.igem.org

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Team Sorbitol:

The 10 colonies were harvested with a Qiagen Miniprep Kit and the DNA was digested with Xba1 and HindIII for 1 hour at 37C.
The digestion was then run on an agarose gel along with controls and undigested vector.
We found that each colony contained accurate bands indicating pBAD30 as well as srlD.
Plated these accurate colonies and stored them all in a fridge at 4C as well as grew some colonies overnight.
Team Fungus:
Purified pET28a.
Made an overnight BL21 culture.
Streaked out BL21 plate.
Ran a PCR with 7/1 PCR product dilute.
Ran out PCR on 1% agarose gel. No products.
Ran PCR (PCR2) using TAQ and only N-limited cDNA.
Digested purified pET28a with EcoRI and XhoI.
Ran out digestion on gel. Inconclusive results again.
Ran a PCR (PCR3) using diluted PCR products from 7/3.