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, LP

• 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.

9/5/13

LP

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

9/6/13

JL, LP

• Purified phage propagation sample from 9.4 via centrifugation (5 minutes at 3000 rpm) and cholorform. Spot tests were then performed to estimate phage titer.

• Took pictures of plates from 8.16 Modeling phage plaque size - Experiment One using alpha imager.

• Divided up assignments for updating notebook.

9/7/13

JL

• Started approximately 15 mL E coli B liquid culture overnight.

• Worked on data analysis / picture processing for 8.16 Modeling phage plaque size - Experiment One.

9/8/13

JL

• Continued data analysis / picture processing for 8.16 Modeling phage plaque size - Experiment One.

• Performed preliminary titer in preparation for repeating the modeling experiment with T1 and T2. For specifics, please see 8.16 Modeling phage plaque size - Experiment One.

• Started approximately 25mL of E coli B liquid culture overnight

9/9/13

JL, LP

• Discussed analysis of modeling result using ImageJ.

• Repeated the modeling procedure in 8.16 Modeling phage plaque size - Experiment One for T1 and T2. Incubation started at 4:30pm.

• Started a fresh round for phage propagation for the Phage Purification Team. 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.

• Divided up assignments for updating the wiki.

9/10/13

JL, LP

• Took the T1 and T2 modeling plates our the incubation at 4:30pm.

• Started approximately 25mL of E coli B liquid culture overnight.

• Measured plaque sizes using ImageJ.

9/11/13

JL, LP

• Isolated the phage (from 9.9) by centrifuging down the bacteria, transferring the supernatant to a new 15mL centrifuge tube, and adding 900ul of chloroform to the supernatant.

• Took pictures of T1 and T2 plates for 8.16 Modeling phage plaque size - Experiment One.

9/12/13

JL

• Spot test showed plaques up to -7. This indicate that the phage suspension solution we gave the Phage Purification Team for CsCl was at least 10E9 pfu/mL.

• Started approximately 30mL of E coli B liquid culture overnight.

9/13/13

JL, LP

• The Phage Purification Team has good news for us. They were able to observe banding in their latest CsCl gradient for T7. We thus decided to perform a new round of mutagenesis this weekend and hopefully selection next week.

• In preparation for mutagenesis tomorrow, we performed titers for 8.24 T7 stock.

• Started approximately 5mL of E coli B liquid culture overnight.

• Streaked out E coli B.

9/14/13

JL

• Autoclaved ddH₂O, LB, x2 top agar.

• Used the autoclaved ddH₂O to make adenine and uracil solution. Specifically,

- 254mg of uracil was dissolved in 10mL of ddH₂O to produce a uracil solution with a concentration of approximately 2.5mg/mL

- 753mg of adenine hemisulfate dihydrate was dissolved in 10mL of ddH₂O to produce a adenine solution with a concentration of approximately 5mg/mL

• Started approximately 5mL of E coli B liquid culture overnight.

9/14/13

JL, LP

• Performed the mutagenesis procedure in 9.13 Mutagen Concentration Test - Ninth Protocol

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

• Started phage propagation