1. Genotyping the intron 22 XbaI A restriction fragment length polymorphism (RFLP) using long distance PCR (LD-PCR).
Genotyping the intron 22 XbaI A restriction fragment length polymorphism (RFLP) using long distance PCR (LD-PCR). A primer specific for factor VIII gene sequence (1A or 2A) is used in conjunction with a primer that flanks the XbaI A site and that hybridises within the int22h homologue (1B or 2B, respectively). The resulting LD-PCR product (∼ 7 kb) contains XbaI A (asterisk) and can be genotyped by XbaI digestion. The analysis is specific for XbaI A because the sequences to which primers 1A or 2A hybridise do not occur at the extragenic int22h loci. Primers 1B or 2B can hybridise, but in the absence of primers 1A and 2A amplification cannot take place.
D J Bowen Mol Path 2002;55:
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