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

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* Started approximately 5mL of E coli B liquid culture overnight.
* Started approximately 5mL of E coli B liquid culture overnight.
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<font size="4"> '''9/14/13''' </font>
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JL, LP
JL, LP

Latest revision as of 21:37, 9 October 2013


Small Phage September - October Notebook: September 1 - September 15 Daily Log



Small Phage
March-April
May-June
July-August
September-October

9/1/13

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


9/2/13


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.
  • Divided up assignments for updating notebook.


9/7/13

JL

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


9/8/13

JL

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


9/9/13

JL, LP

  • Discussed analysis of modeling result using ImageJ.
  • 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.


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.

9/15/13

JL, LP

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