Specific analysis of the intron 22 XbaI polymorphism of the human factor VIII gene using long-distance PCR.
A rapid, non-radioactive, PCR-based method to genotype the XbaI restriction fragment length polymorphism of the human factor VIII gene is described. The method uses long-distance PCR followed by XbaI restriction digestion and agarose gel electrophoresis. The 6.6 kb amplification product includes a constant XbaI site, which provides a digestion control. The specificity of the method was challenged by a blind experiment with 16 genomic DNA samples previously genotyped by Southern blot analysis: a perfect correlation was obtained between genotypes determined using Southern blot and PCR.
De Brasi
C D
CD
Departamento de Genetica, Instituto de Investigaciones Hematologicas Mariano R. Castex, Academia Nacional de Medicina, Buenos Aires, Argentina.
Bowen
D J
DJ
Collins
P W
PW
Larripa
I B
IB
D015139:Blotting, Southern; D004587:Electrophoresis, Agar Gel; D005169:Factor VIII / Q000235:genetics*; D006801:Humans; D007438:Introns / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods; D011110:Polymorphism, Genetic; D012150:Polymorphism, Restriction Fragment Length
D005169:Factor VIII