A semi-automated method for analysis of intron 13 and intron 22 dinucleotide repeat polymorphisms of the factor VIII gene

Clin Lab Haematol. 1996 Jun;18(2):111-4. doi: 10.1046/j.1365-2257.1996.00163.x.

Abstract

A semi-automated method has been developed for the simultaneous analysis of two micro-satellite repeat polymorphisms located in intron 13 and 22 of the Factor VIII gene. The fluorescent dyes, 6-FAM-and HEX-phosphoramidites were used to 5'-end label the respective 5' primers of these two microsatellite repeats and a multiplex polymerase chain reaction (PCR) devised for amplification. The PCR product was loaded onto the gel with DNA size marker labelled with ROX. A total of 24 samples could be analysed simultaneously on an automated DNA sequencer. The results were computed using a dedicated software, with assignment of PCR product size in basepair. This method compares well with the conventional manual procedure using radio-labelled primers, but at the same time overcomes many of the inherent disadvantages associated with the latter method.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alleles
  • Automation
  • DNA Mutational Analysis / methods*
  • DNA Primers
  • Dinucleotide Repeats*
  • Electrophoresis, Polyacrylamide Gel
  • Factor VIII / genetics*
  • Female
  • Fluorescent Dyes
  • Genotype
  • Haplotypes / genetics
  • Hemophilia A / blood
  • Hemophilia A / genetics*
  • Humans
  • Introns / genetics*
  • Leukocytes / chemistry
  • Male
  • Polymerase Chain Reaction / methods*
  • Software

Substances

  • DNA Primers
  • Fluorescent Dyes
  • Factor VIII