Team:BYU Provo/Notebook/Phage Purification/Winterexp/Period1/Exp/6.7 Phage Suspension Buffer Preparation

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Latest revision as of 00:29, 28 September 2013

Phage Purification May - June Notebook: Experiments

Phage Purification

6.7 Phage Suspension Buffer Purification

I) Purpose

Create 1 L of 10X Phage Supension Buffer stock.

II) Expected Outcome

Clean, usable stock of phage suspension buffer.

III) Reagants Used

NaCl

MgCl2

Tris-HCl

IV) Actual Procedure

Put 12.4 g Tris-HCl in a 1 L flask. Added 1 L H2O. Titrated to 7.2 pH using 50% NaOH. (.1 M Tris-HCl)

Added 58.44 g NaCl (1M)

Added 9.52 g MgCl2 (.1M)

Autoclaved and stored in a sealed bottle for use.

V) Results

Created 1 L of 10X phage suspension buffer

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 <font color="#333399" size="3" font face="Calibri">
-: [[Team:BYU_Provo/Small_Phage|Overview]]
+<font size = "4">
+: <u> '''Phage Purification''' </u> </font>
 : [[Team:BYU Provo/Notebook/Phage_Purification/Winterexp|March-April]]
@@ Line 26: / Line 28: @@
 </font>
-| style="width: 82%; background-color: transparent;"|
+| style="width: 80%; background-color: transparent;"|
 <font face="Calibri" size="3">
-<font size="5"> '''6.5 Bacteria Cultures''' </font>
+<font size="5"> '''6.7 Phage Suspension Buffer Purification''' </font>
 <br>
 '''I) Purpose'''
-: Grow bacteria for later use in preparation of phage.
+: Create 1 L of 10X Phage Supension Buffer stock.
 '''II) Expected Outcome'''
-: Viable liquid cultures of both W3110 and BL21
+: Clean, usable stock of phage suspension buffer.
 '''III) Reagants Used'''
-: W3110 and BL21 freezer stock
+: NaCl
-: LB
+: MgCl2
+: Tris-HCl
 '''IV) Actual Procedure'''
-: Infect 45 mL of LB with 2 loop-fulls of bacterial freezer stock and let sit at 37<sup>◦</sup> C for 1 hour.
+: Put 12.4 g Tris-HCl in a 1 L flask. Added 1 L H2O. Titrated to 7.2 pH using 50% NaOH. (.1 M Tris-HCl)
-: Infect 4 mL of LB with 1 mL of liquid culture prepared in previous step.
+: Added 58.44 g NaCl (1M)
-: Let both liquid cultures sit overnight at 37<sup>◦</sup> C.
+: Added 9.52 g MgCl2 (.1M)
+: Autoclaved and stored in a sealed bottle for use.
 '''V) Results'''
-: A
+: Created 1 L of 10X phage suspension buffer
 </font>