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Specific analysis of the intron 22 Xba I polymorphism of the human factor VIII gene using long‐distance PCR
Author(s) -
De Brasi Carlos D.,
Bowen Derrick J.,
Collins Peter W.,
Larripa Irene B.
Publication year - 1999
Publication title -
british journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.907
H-Index - 186
eISSN - 1365-2141
pISSN - 0007-1048
DOI - 10.1046/j.1365-2141.1999.01731.x
Subject(s) - microbiology and biotechnology , agarose gel electrophoresis , southern blot , restriction fragment length polymorphism , restriction digest , genotype , genomic dna , biology , restriction enzyme , polymerase chain reaction , terminal restriction fragment length polymorphism , intron , gene , genetics
A rapid, non‐radioactive, PCR‐based method to genotype the Xba I restriction fragment length polymorphism of the human factor VIII gene is described. The method uses long‐distance PCR followed by Xba I restriction digestion and agarose gel electrophoresis. The 6.6 kb amplification product includes a constant Xba I 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.