GENOTYPING OF THE ABO BLOOD GROUP SYSTEM IN IRAQ POPULATION USING PCR-RFLP

Due to a large number of alleles that could give similar phenotypes but differ in genomic structure. This study study was conducted to determine the ABO genotyping by use PCR-RFLP method. Blood samples withdrawn from 30 unrelated individuals who appear to be healthy. ABO system phenotype grouping was detected by two immunoagglutination tests. Slide and ELISA were done for all samples. Homozygous allele was selected by chosen special family which gives known offspring according to Mendel laws. The DNA was extracted by using two methods, the manual method by using extra gene kit gives relatively high yield concentration (5-10μg/ml) but slightly low purity (0.9-3.3) and the automated BioRobot EZ1 gives slightly lower yield of DNA(4-8 μg/ml) but high purity (1.6–2.2) in comparison with manual method . Two separate segments of ABO gene the glycosyltransferases gene (216pb) in exon 6 and (703pb) in exon 7 were amplified using two sets of specific primers. Restriction digisten of the two amplification PCR productes was used for discrimination of AB and O allells.