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Team:BYU Provo/Notebook/Cholera - Enzyme/July-August/Period3/Dailylog
From 2013.igem.org
| Cholera - Enzymes Notebook: May 1 - May 14 Daily Log
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8/5/13 We got a frozen culture of V. cholerae from Dr. Robison’s lab. Cells are in 20% glycerol. We are waiting for the DspB colony PCR to finish so that we can grow up both the DspB and Savinase at the same time to sequence them and get them ready for protein purification.
8/7/13 The DspB colony PCR didn’t work. We re-ran the PCR yesterday, so we will run the product on gel today, clean up the PCR and sequence it. We also prepped the Savinase for sequencing today since the DspB didn’t work and we don’t want to wait any longer. We also streaked a new plate of cholera with the culture that we received from Dr. Robison’s lab. New plate was placed in the 37° incubator.
8/9/13 Phusion PCR of Savinase and DspB. We ran two samples of each with a control:
8/12/13 We ran DspB and Savinase on gel but the results were not promising. There were two very faint bands for DspB, but not enough for us to use. Savinase did not have any bands showing at all. We then reran the Phusion PCR for our samples following the protocol listed in the Grose lab protocol packet. Our samples were:
8/13/13 We set up PCR again for DspB and Savinase. The PCR tubes were labeled:
8/14/13 We ran the PCR products from yesterday on gel, but nothing showed, not even the ladder. This indicates that when the gel was prepared, the ethidium bromide was not added as it was supposed to be. We made a new gel and re-ran our PCR products:
[IMAGE] There was a faint band for the DspB, but nothing for Savinase. We will try to move forward with our DspB, but may need to restreak the Bacillis subtilis for our Savinase enzyme.
8/21/13 Insert Text
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