Team:BYU Provo/Notebook/SmallPhage/Fallexp/Period1/Dailylog

Small Phage

March-April

May-June

July-August

September-October

• Performed additional spot tests for 12 samples to verify results of phage concentration in each aliquot for 8.26 Mutagen Concentration Test - Eighth Protocol.

• Started approximately 10mL of E coli B liquid culture overnight.

9/2/13

• Worked on compiling data for 8.26 Mutagen Concentration Test - Eighth Protocol; there are significant differences between CsCl gradient for control sample and that for mutated phage sample.

9/3/13

• Started approximately 20 mL of E coli B liquid culture overnight.

9/4/13

JL, LJP

• Discussed results from recent spot test for 8.26 Mutagen Concentration Test - Eighth Protocol. Phage too spread out to effectively select for mutant phage. Decided to stop until the Phage Purification Team can get the wildtype T7 phage to band.

• Discussed plans and designs for team wiki with Darren and Keltzie.

• Started T7 propagation to generate enough phage for the Phage Purification Team to experiment with. Specifically, to a 15mL centrifuge tube we added 8mL of LB, 2mL of E coli B liquid culture overnight, and 20uL of 8.24 T7 phage stock.

8/21/13

• Spot test for T7 only showed plaques up to -5. Might not be high enough for mutagenesis. Need to propagate.

• Plated the S1-S12 and B1-B12 samples in 8.14 Mutagen Concentration Test - Seventh Protocol.

• Started approximately 6 mL of E coli B liquid culture overnight

8/22/13

• Nothing showed on the S1-S12 and B1-B12 plates. Wildtype had too many plaques (T7 at -4).

• T7 propagation for 8.26 Mutagen Concentration Test - Eighth Protocol.

• Started approximately 20 mL of E coli liquid culture overnight.

8/23/13

- Repeated plating of S1-S12 and B1-B12 in 8.14 Mutagen Concentration Test - Seventh Protocol.

8/24/13

- Still no bacteria or phage plaques showed up for S1-S12 and B1-B12 in 8.14 Mutagen Concentration Test - Seventh Protocol -> We decided to focus on 8.26 Mutagen Concentration Test - Eighth Protocol instead

- Purified T7 phage propagation and performed a spot test to estimate the titer of this fresh stock (8.24).

- Started approximately 10mL of E coli B liquid culture overnight

8/25/13

- Titer was performed for T7 phage 8.24 stock at -6 and -7 to gain a more accurate estimate of phage concentration.

- Started approximately 10mL of E coli B liquid culture overnight.

8/26/13

- Performed the actual mutagenesis procedure in 8.26 Mutagen Concentration Test - Eighth Protocol.

- Phage Purification Team was able to run the CsCl gradient right after the completion of mutagenesis.

- Started approximately 20mL of E coli B liquid culture overnight.

8/27/13

- Spot test was performed for the post CsCl aliquots (for both control and mutated group) to estimate phage concentration in each. For specific procedure please refer to 8.26 Mutagen Concentration Test - Eighth Protocol.

8/30/13

- Started approximately 20mL of E coli B liquid culture overnight.

- Worked on editing the abstract for submission.

8/31/13

- Further dilution series and spot test was performed to gain a more accurate idea of phage distribution in CsCl gradient. Specific procedures can be found at 8.26 Mutagen Concentration Test - Eighth Protocol.

- Started approximately 10mL of E coli B liquid culture overnight.