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Table 2 PCR primers and hybridization probes used in estrogen receptor-α genotyping by real-time PCR combined with melting curve analysis

From: Estrogen receptor-α polymorphism in a Taiwanese clinical breast cancer population: a case–control study

Polymorphism site

Melting Temperature (°C)

Oligonucleotide sequences

Exon 1/codon 10

  

   TCT

56

Primers for PCR reaction:

   TCC

64

   Sense: 5'-GGTTTCTGAGCCTTCTGCCCTG-3' (301–322)

  

   Antisense: 5'-AGGCCGGTCTGACCGTAGA-3' (593–575)

  

Hybridization probes:

  

   Sensor: 5'-ACACCAAAGCATCCGGGATGred-3' (377–396)

  

   Anchor: 5'-fluoresceinCCCTACTGCATCAGATCCAAGGGAACG-3' (398–424)

Exon 4/codon 325

  

   CCC

58.5

Primers for PCR reaction:

   CCG

65.5

   Sense: 5'-ACCTGTGTTTTCAGGGATACGA-3' (336–357)

  

   Antisense: 5'-GCTGCGCTTCGCATTCTTAC-3' (705–686)

  

Hybridization probes:

  

   Sensor: 5'-GCTGAGCCCCCGATACTCTAred-3' (554–573)

  

   Anchor: 5'-fluoresceinCCGAGTATGATCCTACCAGACCCTTCA-3' (576–602)

Exon 8/codon 594

  

   ACG

61

Primers for PCR reaction:

   ACA

67

   Sense: 5'-CTGTGTCTTCCCACCTACAG-3' (337–356)

  

   Antisense: 5'-GGGTAAAATGCAGCAGGGATT-3' (641–621)

  

Hybridization probes:

  

   Sensor: 5'-TCCCTGCCACAGTCTGAGAGCred-3' (575–595)

  

   Anchor: 5'-fluoresceinCCCTGGCTCCCACACGGTTCAG-3' (597–618)