Team:BYU Provo/Notebook/SmallPhage/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

- Discussed plans for the selection process of 5.20 Mutagen Concentration Experiment and calculated the needed volume of agar, overnight, and phage

- Made 500mL x8 agar

- Performed dilution series to generate enough 200ug -4 phage stock for selection.

6/6/13

- Started approximately 50mL of BL21 liquid culture overnight.

6/7/13

- Performed the selection process of 5.20 Mutagen Concentration Experiment. Specifically we plated 45 plates of 200ug mutagen, -4 phage dilution using x8 agar. 5 plates of 0ug, -3 phage dilution were also done a control.

5/22/13

- Performed spot test for 5.20 Mutagen Concentration Experiment

- Ran agarose gel to confirm PCR product