Team:BYU Provo/Notebook/Phage Purification/Winterexp/Period11/Dailylog

From 2013.igem.org


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



Phage Purification
March-April
May-June
July-August
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8/19/13

- Performed a cesium chloride gradient on T4. The gradient did not show any band and we aren't sure where the phage are in the gradient. It is possible that we mixed the gradient before or after centrifugation which could have destroyed the band.

8.19 CsCl gradient

AB DL AC


8/21/13

- Discussed and weighed out the T4 cesium chloride gradient for next class. We also discussed a new gradient for T7. We are hoping this gradient will narrow down exactly where the mutant phage will band and where the wild type phage will band. We feel that the phage will differentiate around the 1.3 concentration. By creating a gradient with small steps both above and below 1.3 we should be able to separate the phage out.

AB DL AC


8/23/13

- Prepared a Cesium Chloride Gradient for T4

8.23 CsCl gradient

AB DL AC


8/26/13

- Reviewed results for the T4 gradient. -Ran CsCl Gradient on T7.

8.26 T7 CsCl gradient

We are having issues getting the T7 to band. When we remove the gradient after centrifuging we are unable to see any band at all. A spot test reveals phage are found all over the gradient. We have had this same result several times and believe that the cesium chloride we are using could be the factor. Our plan is to do a very broad test and see if we can get the phage to band anywhere.

AB DL AC


8/28/13

- Discussed the possible gradients we could perform to get T7 to band. We are thinking of running a wild type gradient with different concentrations at 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, and 1.7. This should give us a rough estimate of where the wild type will band.

AB DL AC



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