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Table 1 Primers and conditions for the analysis of FcγRIIIA gene polymorphism

From: Analysis of in vitro ADCC and clinical response to trastuzumab: possible relevance of FcγRIIIA/FcγRIIA gene polymorphisms and HER-2 expression levels on breast cancer cell lines

Namea

Primer sequence

Amplicon size (bp)

Sequence to analyze

FcγRIIIA-F

AAGTCATTTGGGGTCAATTTC

2416d

 

FcγRIIIA-R

CAGAATAGTTTCATCTCGTATATCb

  

FcγRIIIA-R-S

CAGGAATAAGGTGACGGTGG

598

 

Py-FcγRIIIA-F

Bio-AAAGCCACACTCAAAGACAGCc

122e

MAAGCCCCCTGCAGAAGTAGf

Py-FcγRIIIA-R

ATTCCAGGGTGGCACATGTC

  

Py-FcγRIIIA-S

ACACATTTTTACTCCCAA

  
  1. aGenbank reference sequence: NG_009066.1; F forward, R reverse, Py pyrosequencing, S sequence, Bio biotinylated
  2. bIn italic, underlined, is indicated the deliberate mismatch, from the reference sequence, introduced to improve primer stability
  3. cTo increase the specificity of the FcγRIIIA amplification, the pyrosequencing primer was designed to include a cytosine (bold underlined) that is replaced by a thymidine in the FcγRIIIB gene (genebank reference sequence. NG_032926.1)
  4. dCycling conditions: 30″ at 94 °C followed by 35 cycles at 94 °C for 30″, 56 °C for 30″, 68 °C for 2′45″ and a final cycle at 70 °C for 10′
  5. eCycling conditions:10′ at 95 °C followed by 45 cycles at 95 °C for 30″, 57 °C for 30″, 72 °C for 30″ and a final cycle at 70 °C for 10′
  6. fThe polymorphism, in bold, is reported according to the IUB (International Union of Biochemistry) nomenclature