Team:BYU Provo/Notebook/CholeraDetection/Springexp/Period3/Dailylog

5/28/13

- Started three 8mL E coli BL21 liquid culture at around 4pm.

5/29/13

- Continued 5.20 Mutagen Concentration Experiment

- Prepared sample for sequencing. This is done as part of 5.20 T7 Minor Capsid Protein PCR

- Dr. Studier responded to our email today! We also received the E coli stains containing plasmids with clone T7 genes. WE CAN START SITE DIRECTED MUTAGENESIS SOON!

5/30/13

- Plates from yesterday are taken out of incubation at around 4:00pm

5/31/13

- Discussed results for 5.20 Mutagen Concentration Experiment -> will need to try this one more time with minor adjustment to experimental design: plate -4 dilution using x6 and x8 top agar.

- Discussed plans for next week.

- Made new LB and x6 top agar.

6/2/13

- Made about 20ml of BL21 overnight

6/3/13

- Made new LB plates

- Plated -4 titer on x6 and x8 plates for 5.20 Mutagen Concentration Experiment

6/5/13

Regarding our PCR attempt yesterday, we did see a thick fat band ... at the bottom of the gel! Once again our PCR failed. This has led us to propose several reasons why we can't PCR amplify CRO: 1) the Lambda prophage isn't actually in TT 9901 or TT9907, or 2) our primers aren’t functioning to amplify CRO. We streaked a few plates of TT9901 and TT9907 to subject to UV light. If we see plaques, then we will know Lambda has been induced. Meanwhile Dr. Grose is going to check that our primers are the correct ones.