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

From 2013.igem.org

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(Created page with "{{TeamBYUProvo}} <br> {| width="100%" | colspan="3" | <font color="#333399" size="5" font face="Calibri"> '''Small Phage May - June Notebook: May 1 - May 12 Daily Log'''</font>...")
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{| width="100%"
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| colspan="3" | <font color="#333399" size="5" font face="Calibri"> '''Small Phage May - June Notebook: May 1 - May 12 Daily Log'''</font>
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| colspan="3" | <font color="#333399" size="5" font face="Calibri">  
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: '''Phage Purification July - August Notebook: August 17 - August 31 Daily Log'''</font>
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: [[Team:BYU_Provo/Small_Phage|Overview]]
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: [[Team:BYU_Provo/Phage_Purification|Overview]]
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: [[Team:BYU Provo/Notebook/SmallPhage/Winterexp|March-April]]
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: [[Team:BYU Provo/Notebook/Phage_Purification/Winterexp|March-April]]
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: [[Team:BYU Provo/Notebook/SmallPhage/Springexp|May-June]]
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: [[Team:BYU Provo/Notebook/Phage_Purification/Springexp|May-June]]
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: [[Team:BYU Provo/Notebook/SmallPhage/Summerexp|July-August]]
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: [[Team:BYU Provo/Notebook/Phage_Purification/Summerexp|July-August]]
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: [[Team:BYU Provo/Notebook/SmallPhage/Fallexp|September-October]]
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: [[Team:BYU Provo/Notebook/Phage_Purification/Fallexp|September-October]]
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<font size="4"> '''5/1/13''' </font>
 
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- Commencement of spring!!!
 
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- Discussed goals and outlined plans for spring term
 
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<html> <a href="https://2013.igem.org/Team:BYU_Provo/Notebook/Phage_Purification/Winterexp/Period10/Dailylog"><< Previous</a>
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<font size="4"> '''5/2/13''' </font>
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<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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- Designed primers for amplifying and sequencing phage capsid protein
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- Wrote emails inquiring mutagen (NG) and plasmid for in vitro assembly
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<font size="4"> '''5/3/13''' </font>
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<font size="4"> '''8/7/13''' </font>
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- Performed agar test, focusing primarily on ×8
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- Performed a cesium chloride gradient purification on T7 mutated phage from the small phage team.
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: [[Team:BYU_Provo/Notebook/SmallPhage/Springexp/Period1/Exp/Team:BYU_Provo/5.3 T7 phage selection method test|5.3 T7 phage selection method test]]
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:[[Team:BYU_Provo/Notebook/Phage_Purification/Winterexp/Period1/Exp/8.7CsClGradient|8.7 CsCl Gradient]]
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- Processed phage amplification
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: [[Team:BYU_Provo/Notebook/SmallPhage/Springexp/Period1/Exp/Team:BYU_Provo/5.3 Phage Amplification/Purification|5.3 T7 phage amplification/purification]]
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<font size="4"> '''5/4/13''' </font>
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<font size="4"> '''8/9/13''' </font>
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- Performed dilution series using stock 5.3 (-1 through -11)
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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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- Started two 5mL overnights of BL21
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<font size="4"> '''5/5/13''' </font>
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<font size="4"> '''8/12/13''' </font>
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- Spot test using stock 5.3 and its dilution series (from 5.4)
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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/SmallPhage/Springexp/Period1/Exp/Team:BYU_Provo/5.3 Phage Amplification/Purification|5.3 T7 phage amplification/purification]]
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- Started liquid culture for purification team (at around noon)
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: 1mL of BL21 overnight + 4mL of LB + 100μL -2 phage (received stock)
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- Started 5.5 amplification from a plaque test
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: [[Team:BYU_Provo/Notebook/SmallPhage/Springexp/Period1/Exp/Team:BYU_Provo/5.5_Amplification_from_a_plaque_test|5.5 Amplification from a plaque test]]
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<font size="4"> '''5/6/13''' </font>
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</font>
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- Continued [[Team:BYU_Provo/Notebook/SmallPhage/Springexp/Period1/Exp/Team:BYU_Provo/5.3 T7 phage selection method test|5.3 T7 phage amplification/purification]]
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<html> <a href="https://2013.igem.org/Team:BYU_Provo/Notebook/Phage_Purification/Winterexp/Period10/Dailylog"><< Previous</a>
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- Performed liquid culture phage concentration test
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<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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: [[Team:BYU Provo/Notebook/SmallPhage/Springexp/Period1/Exp/Team:BYU Provo/5.6 T7+ Liquid Culture Phage Concentration Test|5.6 T7+ Liquid Culture Phage Concentration Test]]
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<font size="4"> '''5/7/13''' </font>
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- Started two 5mL of E coli BL21 overnight
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- Designed procedure for applying mutagen and selecting for T7
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<font size="4"> '''5/8/13''' </font>
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- Went over procedure for applying mutagen and PCR with Dr. Grose
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- Performed spot tests under [[Team:BYU Provo/Notebook/SmallPhage/Springexp/Period1/Exp/Team:BYU Provo/5.6 T7+ Liquid Culture Phage Concentration Test|5.6 T7+ Liquid Culture Phage Concentration Test]]
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- Started two 5mL BL21 overnights
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- Learned how to create LB plates
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<font size="4"> '''5/9/13''' </font>
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- Practiced with ×6 and ×8 top agar
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: [[Team:BYU_Provo/Notebook/SmallPhage/Springexp/Period1/Exp/5.9_T7_selection_method_test|5.9 T7 Selection Method Test #2]]
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- Started [[Team:BYU Provo/Notebook/SmallPhage/Springexp/Period1/Exp/5.9 T7+ Liquid Culture Phage Concentration Test|5.9 T7+ Liquid Culture Phage Concentration Test #2]]
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- Discussed plans and schedule for next week.
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<font size="4"> '''5/10/13''' </font>
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- Worked on [[Team:BYU_Provo/Notebook/SmallPhage/Springexp/Period1/PR|Progress Report]]
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<font size="4"> '''5/12/13''' </font>
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- Started two 5mL overnight at around 6pm
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{{TeamBYUProvoFooter}}
{{TeamBYUProvoFooter}}

Revision as of 22:15, 4 September 2013


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



Overview
March-April
May-June
July-August
September-October


<< Previous Next >>


8/7/13

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


<< Previous Next >>

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