Unformatted text preview: them to the side of the vessel, after placing near a magnet for 30–60 s. Purification procedures may even use crude cell extracts for binding of 6xHis-tagged protein. 2. Expression systems are used to produce copies of a desired protein within a host cell. T7 RNA polymerase and the lac promoter. Lactose. Steps: Get a column, white lid and yellow bottom lid. Pack a 1mL to 1.5 Ni-NTA Column. Don’t need to use glass rod to pack affinity column. Binding capacity of Ni-NTA is 5 to 10mg proteins per 1 mL resin so 1 column is enough per group. Wash the column with 5 column column dH2O, followed by 5 column volume of buffer 1. Cap the bottom lid, leave 2mL buffer 1 above the top bed of the resin. Label your column with a tape, instead of writing on the column. Cap the top lid tight. Wrap the white cap with Para-film. Then put your column upward in a big beaker and save it in the cold room....
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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.
- Spring '11