Freezing the single hair bulb in the liquid nitrogen,heating to 94℃ f0r three times and directly using in PCR, we successfully typed the variable number of tandem repeats(VNTR) in the intron 40of the von Willebrand factor(vWF) gene. 62 hair bulbs out of 68 freezed hair samples(91. 2% ) weresuccessfully amplifled and typed.
Objective:The VNTR in the intron40 of vWF gene was amplified by direct PCR and two amplified fragments from heterozygous persons were isolated by a high-resolution polyacrylamide gel electrophoresis. After elution from gel,the preliminary products were amplified again,and the amplified fragments were digested with Alu I,then the genotype could be directly determined through eletrophoresis.
This result showed that the VNTR in the intron40 of vWF gene remained highly genetic polymorphism in Guangdong Han population and the locus might be useful as a genetic marker for paternity identification in Guangdong population.
Methods The hF VIll cDNA (β domain deleted)was first inserted into pAdCMV - linker 1 to become a vector plasmid pAdI - hFVIII with SV4Ointron between CMV promotor and hFVIII cDNA fragment.
We shall report a new method for the detection of mixed chimerism, which makes use of the fact, that the von Willebrand factor (vWF) gene has a highly variable region-variable number of tandem repeats (VNTR) - within intron 40.
Two markers (vWF 1 and vWF 2) were used in the polymorphic region of ATCT repeats (variable number of tandem repeat, VNTR) within intron 40 of the vWF gene.