Skip to main content

Table 3 Primers used for the generation and verification of yeast single knockouts

From: ORMDL proteins are a conserved new family of endoplasmic reticulum membrane proteins

Oligonucleotides for the synthesis of kanMX4 cassettes  
Primer Sequence 5' to 3'
   YGR038w F AGG AAC TGC TGA GGC GGC TTC TAC CTC GTA TAG TCG AAA TCA AAC CGT ACG CTG CAG GTC GAC
   YGR038w R TGT TCA ACT AAT TTG GGC GCG ACC TGT GAT ACC TGG GAT AGA AAT ATC GAT GAA TTC GAG CTC G
   YLR350w F ATC ATG ATT GAC CGC ACT AAA AAC GAA TCT CCA GCT TTT GCG TAC GCT GCA GGT CGA C
   YLR350w R CTA ACT AAT TTG AGC ACG GCC CGT AAT ACC AGG GAT GGA TAT CGA TGA ATT CGA GCT CG
Oligonucleotides for gene integrity and gene deletion verification
Verification of ORM1 gene integrity  
   A1 (F) GTT TAG GAA CGG ATT ATA GA
   A2 (R) CTG GTC ATT TTC ATC TTC AA
Verification of ORM1 substitution  
   A1 (F) GTT TAG GAA CGG ATT ATA GA
   KAN5 (R) GTT CGG ATG TGA TGT GAG
Verification of ORM2 gene integrity  
   YLR350w up400 (F) GCG TAT TTT GAT TGC TCA TA
   YLR350w ver (R) TGG TTT CAG GTT AGA CAC AT
Verification of ORM2 substitution  
   YLR350w up400 (F) GCG TAT TTT GAT TGC TCA TA
   KAN5 (R) GTT CGG ATG TGA TGT GAG
  1. Oligonucleotides in bold are 18-19 nucleotide homology regions to pFA6a MCS sequences.