CaYPD1 purification-2Pre Lab

CaYPD1 purification-2Pre Lab - Pre Lab CaYPD1...

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Pre Lab CaYPD1 purification-2 Purpose: Purification of dry cell pellet of His6-CaYPD1 or CaYPD1-His6. Use of sonication procedure to break the BL21(DE)3 cell that has expressed the protein of interest CaYPD1. Techniques: Wear ear protection devices to prevent ear damage. Set up a warning sign out of the sonication room (the cold room). Keep your sample on a ice-water bath constantly during the process. If bubbles are generated during sonifcation in your sample, turn it off immediately. All of the tips and eppendorf tubes that are in contact with cells should be disposed into a plastic bag inside of the “autoclavable waste” box. If cells are spilled out, use 75% ethanol to clean the spill. Also use 75% ethanol to clean the autoclave waste beaker. For nonautoclave waste, dispose it in the trash can. Steps: Obtain a tube of dry cell pellet and keep it on ice. Add 1 mL Buffer 1 to the cell pellet. Dissolve cell pellet. Transfer the dissolved cell to a 2 mL Eppendorf tube. Prepare two eppendorf tubes. Label one as: T (total) followed by plasmid name followed by cell
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This note was uploaded on 10/25/2011 for the course ALL 101 taught by Professor All during the Spring '11 term at Cameron University.

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