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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: 18%; background-color: transparent;"| <font color="#333399" size="3" font face="Calibri"> : [[Team:BYU_Provo/Phage_Purification|Overview]] : [[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: 82%; background-color: transparent;"| <font face="Calibri" size="3"> <font size="5"> '''8.16 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 : phage suspension buffer '''IV) Actual Procedure''' : Create different concentrations of CsCl solutions to create a gradient. :: Add 0.5497 g of CsCl to 2 mL of phage suspension buffer to create a 1.2 g/ml density gradient. :: Add 1.6175 g of CsCl to 4 mL of phage suspension buffer to create a 1.2960 g/ml density gradient. :: Add 1.6207 g of CsCl to 4 mL of phage suspension buffer to create a 1.2966 g/ml density gradient. :: Add 0.8128 g of CsCl to 2 mL of phage suspension buffer to create a 1.2975 g/ml density gradient. :: Add 1.6288 g of CsCl to 4 mL of phage suspension buffer to create a 1.2981 g/ml density gradient. :: Add 1.6332 g of CsCl to 4 mL of phage suspension buffer to create a 1.2989 g/ml density gradient. :: Add 1.6375 g of CsCl to 4 mL of phage suspension buffer to create a 1.2997 g/ml density gradient. :: Add 1.6408 g of CsCl to 4 mL of phage suspension buffer to create a 1.3003 g/ml density gradient. :: Add 1.6451 g of CsCl to 4 mL of phage suspension buffer to create a 1.3011 g/ml density gradient. :: Add 1.6494 g of CsCl to 4 mL of phage suspension buffer to create a 1.3019 g/ml density gradient. :: Add 1.6527 g of CsCl to 4 mL of phage suspension buffer to create a 1.3025 g/ml density gradient. :: Add 0.8288 g of CsCl to 2 mL of phage suspension buffer to create a 1.3034 g/ml density gradient. :: Add 1.6608 g of CsCl to 4 mL of phage suspension buffer to create a 1.3040 g/ml density gradient. :: Add 1.6407 g of CsCl to 4 mL of phage suspension buffer to create a 1.6 g/ml density gradient. : 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. :Add 4 mL of mutant phage to the top of the gradient in both tubes : 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 at the bottom of the tube. : Extract about 2 mL of phage at a time. Add about 1.5 to one eppendorf. : When the needle is removed place another eppendorf under the hole to catch any leaking phage and fill it with the remainder of the phage in the needle. '''V) Results''' : The phage didn't band anywhere. After running a spot test it appears that the phage where present in every aliquot extracted. [[File:Phage816_1.jpg | thumb|none|alt=]] [[File:Phage816_2.jpg | thumb|none|alt=]] [[File:Phage816_3.jpg | thumb|none|alt=]] </font> |} {{TeamBYUProvoFooter}}

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