Team:BYU Provo/Notebook/Phage Purification/Summerexp/Period1/Exp/7.26 Phage Titer

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: <u> '''Phage Purification''' </u> </font>
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'''III) Reagants Used'''
'''III) Reagants Used'''
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: W3110 freezer stock
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: E Coli B freezer stock
: LB
: LB
: 1.5x top agar
: 1.5x top agar
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'''IV) Actual Procedure'''
'''IV) Actual Procedure'''
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: Infect 500 µL of LB with 1 loop-full of w3110 freezer stock and put on shaker at 37<sup>◦</sup> C for 5 hours.
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: Infect 500 µL of LB with 1 loop-full of E. Coli B freezer stock and put on shaker at 37<sup>◦</sup> C for 5 hours.
: Plate w3110  on an LB plate with 4.5 mL top agar.  
: Plate w3110  on an LB plate with 4.5 mL top agar.  
: Dilute T4 using a dilution series.  
: Dilute T4 using a dilution series.  
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: The titer of T4 phage showed plaques down to a -5 concentration.  This concentration will allow us to perform and EM and examine how pure the phage solution is.
: The titer of T4 phage showed plaques down to a -5 concentration.  This concentration will allow us to perform and EM and examine how pure the phage solution is.
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[[File:Phage_Titer_6.26.13.jpg| thumb|none|alt=A phage plaques were found but with contamination.|T7 Titer]]
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[[File:T4_Phage_Titer_20130729.jpg| thumb|none|alt=A phage plaques were found down to -5.|T4 Titer]]

Latest revision as of 00:35, 28 September 2013


Phage Purification July - August Notebook: Experiments



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

7.26 Titer


I) Purpose

Test the concentration of purified T4 phage.

II) Expected Outcome

Plaques from the bands in the CsCl gradient where phage were purified. Expecting plaques down to at least -5 concentration

III) Reagants Used

E Coli B freezer stock
LB
1.5x top agar
T4 phage from CsCl gradient


IV) Actual Procedure

Infect 500 µL of LB with 1 loop-full of E. Coli B freezer stock and put on shaker at 37◦ C for 5 hours.
Plate w3110 on an LB plate with 4.5 mL top agar.
Dilute T4 using a dilution series.
Spot 2 µL of phage from dilution series onto plate using the concentrations 0, -2, -3, -4, -5, and -6.
Place in 37◦ C incubator overnight.

V) Results

The titer of T4 phage showed plaques down to a -5 concentration. This concentration will allow us to perform and EM and examine how pure the phage solution is.
A phage plaques were found down to -5.
T4 Titer


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