Team:BYU Provo/Notebook/Phage Purification/Winterexp/Period10/Dailylog

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: [[Team:BYU_Provo/Small_Phage|Overview]]
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: [[Team:BYU Provo/Notebook/SmallPhage/Winterexp|March-April]]
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: <u> '''Phage Purification''' </u> </font>
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: [[Team:BYU Provo/Notebook/SmallPhage/Springexp|May-June]]
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: [[Team:BYU Provo/Notebook/Phage_Purification/Winterexp|March-April]]
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: [[Team:BYU Provo/Notebook/SmallPhage/Summerexp|July-August]]
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: [[Team:BYU Provo/Notebook/Phage_Purification/Springexp|May-June]]
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: [[Team:BYU Provo/Notebook/SmallPhage/Fallexp|September-October]]
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: [[Team:BYU Provo/Notebook/Phage_Purification/Summerexp|July-August]]
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: [[Team:BYU Provo/Notebook/Phage_Purification/Fallexp|September-October]]
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<font size="4"> '''8/1/13''' </font>
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- Performed the spot test in [[Team:BYU Provo/Notebook/SmallPhage/Springexp/Period3/Exp/7.29 Mutagen Concentration Test - Sixth Protocol|7.29 Mutagen Concentration Test - Sixth Protocol]].
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<font size="4"> '''8/2/13''' </font>
<font size="4"> '''8/2/13''' </font>
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- Because the top agar used in yesterday's spot was contaminated, we decided to redo this step. Also, because most of the spot tests went down to -6, we performed further dilution for the next testing to get a more accurate idea of phage concentration.
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- Prepared Phage Suspension Buffer
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- Discussed modeling options: model phage plaque size against various variables including phage particle size, agar concentration, and bacterial concentration.
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<br>
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- Streaked E coli W3110 and K12 from frozen stock in preparation for the viability test in [[Team:BYU Provo/Notebook/SmallPhage/Summerexp/8.2 Modeling phage plaque size|8.2 Modeling Phage Plaque Sizes - Experiment 1]].
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<font size="4"> '''8/5/13''' </font>
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- Prepared Cesium Chloride samples for layering on Wednesday
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<font size="4"> '''8/3/13''' </font>
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<font size="4"> '''8/7/13''' </font>
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- E coli W3110 streak worked, but K12 did not survive. Thus, we tried to amplify K12 using liquid culture. For specifics, please consult [[Team:BYU Provo/Notebook/SmallPhage/Summerexp/8.2 Modeling phage plaque size|8.2 Modeling Phage Plaque Sizes - Experiment 1]].
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- Performed a cesium chloride gradient purification on T7 mutated phage from the large phage team.
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:[[Team:BYU_Provo/Notebook/Phage_Purification/Winterexp/Period1/Exp/8.7CsClGradient|8.7 CsCl Gradient]]
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<font size="4"> '''8/4/13''' </font>
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<font size="4"> '''8/9/13''' </font>
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- Started E coli liquid culture over night
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- We performed dialysis on T7 mutated phage from the cesium chloride gradient on 8.7. Because the gradient only showed one band, we decided to not do an EM as we most likely had mutant phage mixed in with wild type. We discussed a gradient to run in the future that will be able to further identify where the phage band.  We decide on a CsCl gradient of concentrations 1.2, 1.25, 1.3, 1.35, 1.4, 1.45, 1.5, 1.6.  We hope to be able to identify exactly where the phage bands so that we can make small variations in the gradient at that point.  This will allow us to separate mutant phage from wild type.
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* 10mL of BL21 for spot test in [[Team:BYU Provo/Notebook/SmallPhage/Springexp/Period3/Exp/7.29 Mutagen Concentration Test - Sixth Protocol|7.29 Mutagen Concentration Test - Sixth Protocol]].
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* 2mL each of BL21, W3110, and B for phage viability and infection test in [[Team:BYU Provo/Notebook/SmallPhage/Summerexp/8.2 Modeling phage plaque size|8.2 Modeling Phage Plaque Sizes - Experiment 1]].
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<font size="4"> '''8/5/13''' </font>
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<font size="4"> '''8/12/13''' </font>
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- Performed Phage viability/infection test for [[Team:BYU Provo/Notebook/SmallPhage/Summerexp/8.2 Modeling phage plaque size|8.2 Modeling Phage Plaque Sizes - Experiment 1]]  
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- Performed a CsCl gradient on the T7 mutant phage from the small phage team.  We used the same phage that we used on 8.7.
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:[[Team:BYU_Provo/Notebook/Phage_Purification/Winterexp/Period1/Exp/8.12CsClGradient|8.12 CsCl Gradient]]
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<font size="4"> '''8/14/13''' </font>
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- We performed dialysis on the phage from [[Team:BYU_Provo/Notebook/Phage_Purification/Winterexp/Period1/Exp/8.12CsClGradient|8.12 CsCl Gradient]]
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:-We also ran another T4 gradient  [[Team:BYU_Provo/Notebook/Phage_Purification/Winterexp/Period1/Exp/8.14 T4 CsCl Gradient|8.14 T4 CsCl Gradient]]
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<font size="4"> '''8/16/13''' </font>
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- Prepared Cesium Chloride samples for centrifugation of T4
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[[Team:BYU_Provo/Notebook/Phage_Purification/Winterexp/Period1/Exp/8.16 T7 CsCl Gradient|8.16 T7 CsCl Gradient]]
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Latest revision as of 00:05, 28 September 2013


Phage Purification July - August Notebook: August 1 - August 16 Daily Log



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



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8/2/13

- Prepared Phage Suspension Buffer



8/5/13

- Prepared Cesium Chloride samples for layering on Wednesday


8/7/13

- Performed a cesium chloride gradient purification on T7 mutated phage from the large phage team.

8.7 CsCl Gradient


8/9/13

- We performed dialysis on T7 mutated phage from the cesium chloride gradient on 8.7. Because the gradient only showed one band, we decided to not do an EM as we most likely had mutant phage mixed in with wild type. We discussed a gradient to run in the future that will be able to further identify where the phage band. We decide on a CsCl gradient of concentrations 1.2, 1.25, 1.3, 1.35, 1.4, 1.45, 1.5, 1.6. We hope to be able to identify exactly where the phage bands so that we can make small variations in the gradient at that point. This will allow us to separate mutant phage from wild type.


8/12/13

- Performed a CsCl gradient on the T7 mutant phage from the small phage team. We used the same phage that we used on 8.7.

8.12 CsCl Gradient


8/14/13

- We performed dialysis on the phage from 8.12 CsCl Gradient

-We also ran another T4 gradient 8.14 T4 CsCl Gradient


8/16/13

- Prepared Cesium Chloride samples for centrifugation of T4

8.16 T7 CsCl Gradient

<< Previous Next >>

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