Team:BYU Provo/Notebook/SmallPhage/Summerexp/Period4/Dailylog

From 2013.igem.org


Small Phage July - August Notebook: August 17 - August 31 Daily Log



Small Phage
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May-June
July-August
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8/17/13

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


8/18/13

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


8/19/13

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


8/20/13

  • Selected for 12 small plaques (S1-S12, relatively large phage) and 12 big plaques (B1-B12, relatively small phage) at respective end of spectrum for 8.14 Mutagen Concentration Test - Seventh Protocol. Spectrum ends were chosen because the likelihood of contamination between different layers of CsCl during the extraction process.
  • Performed dilution series and spot test for T1, T2 (from stock), and T7.
  • Started approximately 30 mL of E coli B liquid culture overnight.


8/21/13

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


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