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Sequence of plasmid/ORF junctions:
5' C TAT CTA TTC GAT GAT GAA GAT ACC CCA CCA AAC CCA AAA AAA GAG ATC GAA TTC CAG CTG ACC ACC ATG XXX XXX XXX XXX XXX XXX *** CAT GGC AAT TCC CGG GGA TCC GTC GAC CTG CAG AGA TCT ATG AAT CGT AGA TAC TGA AAA ACC CCG CAA G 3' Where ATG represents the actual start codon of the yeast ORF. XXX's represent the ORF-specific sequences. ***'s represent one of the 3 stop codons. Primers & Re-PCR primers used for amplifying yeast ORFs and their "common tails": Forward primer: 5' AATTCCAGCTGACCACCATG 3' Reverse primer: 5' GATCCCCGGGAATTGCCATG 3' Re-PCR Forward primer: 5' CTATCTATTCGATGATGAAGATACCCCACCAAACCCAAAAAAAGAGATCGAATTCCAGCTGACCACCATG 3' Re-PCR Reverse primer: 5' CTTGCGGGGTTTTTCAGTATCTACGATTCATAGATCTCTGCAGGTCGACGGATCCCCGGGAATTGCCATG 3' Return to Protocols Page |
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HHMI, |