Human BRCA2 cDNA was cloned into phCMV1 with two repeats of (MBP) at the N-terminus. 293TD cells were transfected and harvested 31 hours post-transfection. Extracts were batch bound to Amylose resin. The protein was eluted with 10 mM maltose, loaded onto HiTrap Q, and step eluted at 450 mM NaCl.
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Human BRCA2 cDNA Purification Protocol
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Other organizations : University of California, Davis
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Variable analysis
independent variables
- Cloning of human BRCA2 cDNA into phCMV1 with two repeats of (MBP) at the N-terminus
- Protein expression and purification
- Transfection of 293TD cells
- Harvesting of cells 31 hours post-transfection
- Batch binding to Amylose resin
- Elution with 10 mM maltose
- Loading onto HiTrap Q column
- Step elution at 450 mM NaCl
- No positive or negative controls were explicitly mentioned in the information provided.
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