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

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Latest revision as of 00:43, 28 September 2013

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

Phage Purification

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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-: [[Team:BYU_Provo/Phage_Purification|Overview]]
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+: <u> '''Phage Purification''' </u> </font>
 : [[Team:BYU Provo/Notebook/Phage_Purification/Winterexp|March-April]]
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-<html>							<a href="https://2013.igem.org/Team:BYU_Provo/Notebook/Phage_Purification/Winterexp/Period10/Dailylog"><< Previous</a>
+<html>													<a href="https://2013.igem.org/Team:BYU_Provo/Notebook/Phage_Purification/Winterexp/Period10/Dailylog"><< Previous</a>
 							<a href="https://2013.igem.org/Team:BYU_Provo/Notebook/Phage_Purification/Fallexp/Period1/Dailylog" style="display:block;float:right;">Next >></a>
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 <br>
-<font size="4"> '''8/7/13''' </font>
+<font size="4"> '''8/19/13''' </font>
-- Performed a cesium chloride gradient purification on T7 mutated phage from the small phage team.
+- 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.
+:[[Team:BYU_Provo/Notebook/Phage_Purification/Winterexp/Period1/Exp/8.19CsClGradient| 8.19 CsCl gradient]]
-:[[Team:BYU_Provo/Notebook/Phage_Purification/Winterexp/Period1/Exp/8.7CsClGradient|8.7 CsCl Gradient]]
+AB DL AC
 <br>
-<font size="4"> '''8/9/13''' </font>
+<font size="4"> '''8/21/13''' </font>
-- 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.
+- 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
 <br>
-<font size="4"> '''8/12/13''' </font>
+<font size="4"> '''8/23/13''' </font>
-- 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.
+- Prepared a Cesium Chloride Gradient for T4
+:[[Team:BYU_Provo/Notebook/Phage_Purification/Winterexp/Period1/Exp/8.23CsClGradient| 8.23 CsCl gradient]]
-:[[Team:BYU_Provo/Notebook/Phage_Purification/Winterexp/Period1/Exp/8.12CsClGradient|8.12 CsCl Gradient]]
+AB DL AC
 <br>
+<font size="4"> '''8/26/13''' </font>
+- Reviewed results for the T4 gradient.
+-Ran CsCl Gradient on T7.
+:[[Team:BYU_Provo/Notebook/Phage_Purification/Winterexp/Period1/Exp/8.26 CsCl Gradient| 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
+<br>
+<font size="4"> '''8/28/13''' </font>
+- 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
+<br>
 </font>