Lecture07_handout - 106 amplification Repeat ~25-30 times...

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Unformatted text preview: 106 amplification Repeat ~25-30 times PCR PCR amplify if needed gen Whole ome GeneX::affinity tag sequence Introduce/express ProteinX-affinity tag Pull-down/co-purify ProteinX+binding partners Mass spec Identify "TAP" (tandem affinity purification)-tag TEV Protein A Calmodulin Gene X site Peptide column + Ca2+ (binds Calmodulin) EDTA elution IgG column (binds protein-A) TEV protease cleavage (gentle!) Elutes ProteinX and any partners ...
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This note was uploaded on 05/07/2008 for the course MCB 130 taught by Professor Schekman during the Spring '08 term at University of California, Berkeley.

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Lecture07_handout - 106 amplification Repeat ~25-30 times...

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