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Primers used for amplifying yeast ORFs
and their "common tails":

Forward primer:

5' AA TTC CAG CTG ACC ACC ATG xxx xxx xxx xxx xxx xxx xxx 3'

whereas the ATG represents the actual start codon of the yeast ORF.

 

Reverse primer

5' xxx xxx xxx xxx xxx xxx *** CAT GGC AAT TCC CGG GGA TC 3' (coding strand)

3' xxx xxx xxx xxx xxx xxx *** GTA CCG TTA AGG GCC CCT AG 5' (actual sequence of primer)

 

whereas the xxx's represent the ORF-specific sequences and the ***'s one of the 3 stop codons.

 

For the re-PCRs the following primers have been used:

Forward primer:

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 3'

Reverse primer:

5' CATGGCAATTCCCGGGGATCCGTCGACCTGCAGAGATCTATGAATC
GTAGATACTGAAAAACCCCGCAAG 3' (coding strand)

3' GTACCGTTAAGGGCCCCTAGGCAGCTGGACGTCTCTAGATACTTAG
CATCTATGACTTTTTGGGGCGTTC 5' (actual primer)

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