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.

6/9/13

- Started 21mL of BL21 overnight.

6/10/13

- Made 1250mL of x8 top agar

- Attempted to amplify phage from larger plaques in 5.20 Mutagen Concentration Experiment

6/11/13

- Started approximately 70mL of BL21 liquid culture overnight

6/12/13

- Perfected protocol for applying mutagen to phage

- Started 6.12 Mutagen Concentration Test - Perfected Protocol

6/13/13

- Started approximately 30mL of BL21 liquid culture overnight.

6/14/13

- Titered the mutagenesis product from Wednesday as part of 6.12 Mutagen Concentration Test - Perfected Protocol

6/16/13

- Started approximately 100mL of BL21 liquid culture overnight.

6/17/13

- Plated 5 controls and 50 selection plates (500ug at -2) as part of selection 1 in 6.12 Mutagen Concentration Test - Perfected Protocol