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PCR primers for KO, tagging, detection amplification of pRS400a (KanMX) or pRS403 (HIS3) for KO
 

Summary: PCR primers for KO, tagging, detection
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amplification of pRS400a (KanMX) or pRS403 (HIS3) for KO:
5'- X40 5' end YFG- GAT TGT ACT GAG AGT GCA CC -3'
5'-X40 3' end YFG- CTG TGC GGT ATT TCA CAC CG -3'
-> ~1.6 kb KanMX PCR product, ~2.1 kb HIS3 PCR product
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KanMX (Reverse Primer) internal detection oligo (use with YFG 3' UTR oligo):
5'- TAC GGG CGA CAG TCA CAT CAT G -3'
KanC3 (Forward Primer) internal detection oligo (use with YFG 5' UTR oligo):
5'- CCT CGA CAT CAT CTG CCC AGA T-3'
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HIS3 internal detection oligo (sense strand; use with YFG 3' UTR oligo):
5'- AGT GCG TTC AAG GCT CTT GC -3'
HIS3 detection Forward oligo (sense strand; use with YFG 3' UTR oligo):
5'- CGA GGC AAG AAT GAT CAT CAC C-3'
HIS3 detection Reverse oligo (antisense strand; use with YFG 5' UTR oligo?):
5'- GGG AAG ATC GAG TGC TCT ATC G-3'
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amplification of pAG25 (natMX) for KO:

  

Source: Auble, David - Department of Biochemistry and Molecular Genetics, University of Virginia

 

Collections: Biology and Medicine