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{{TeamBYUProvo}} <br> {| width="100%" | colspan="3" | <font color="#333399" size="5" font face="Calibri"> '''Phage Purification July - August Notebook: Experiments'''</font> <br> <br> |- valign="top" | style="width: 20%; background-color: transparent;"| <font color="#333399" size="3" font face="Calibri"> <font size = "4"> : <u> '''Phage Purification''' </u> </font> : [[Team:BYU Provo/Notebook/Phage_Purification/Winterexp|March-April]] : [[Team:BYU Provo/Notebook/Phage_Purification/Springexp|May-June]] : [[Team:BYU Provo/Notebook/Phage_Purification/Summerexp|July-August]] : [[Team:BYU Provo/Notebook/Phage_Purification/Fallexp|September-October]] </font> | style="width: 80%; background-color: transparent;"| <font face="Calibri" size="3"> <font size="5"> '''8.12 CsCl Gradient''' </font> <br> '''I) Purpose''' : Further purify the phage to a high level of purification. '''II) Expected Outcome''' : Purified and viable phage will be extracted from the CsCl gradient. '''III) Reagants Used''' : T7 mutant phage : CsCl : dialysis tubing : phage suspension buffer '''IV) Actual Procedure''' : Create different concentrations of CsCl solutions to create a gradient. :: 1.1 - 2 mL ::1.2 - 2 mL ::1.25 - 2 mL ::1.3 - 2 mL ::1.35 - 2mL ::1.4 - 2 mL ::1.5 - 3 mL ::1.6 - 2 mL : Layer the gradient into two centrifuge tubes, using half of the total volume created for each tube. For example, add 2 mL of 1.3 density to one tube and the other 2 mL to another tube. : Fill the remaining space in the tube with phage suspension buffer to the top. : Centrifuge at 26500 rpms (100,000 g) for 2.5 hours. : Extract using a needle and puncturing the side of the tube, placing the needle underneath the band. : Place phage in dialysis tubing and place in flask with 1 L of phage suspension buffer for 30 minutes at 4^◦ C. : Repeat previous step three more times. : Remove purified phage from dialysis tubing and store in 4^◦ C. '''V) Results''' : The Phage did not band anywhere [[File:Phage12_1.jpg | thumb|none|alt=]] [[File:Phage12_2.jpg | thumb|none|alt=]] [[File:Phage12_3.jpg | thumb|none|alt=]] </font> |} {{TeamBYUProvoFooter}}

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