Genotyping Of ABO gene In Iraqi Population Sample By Using PCR-RFLP Technique

Abstract

The aim of this study was the molecular detection of ABO gene and identify the patterns of alleles in Iraqi population sample. The study consisted of 100 unrelated individuals who appear to be healthy. The blood samples were used for detecting of blood groups and genotyping of ABO gene allele by using PCR-RFLP technique. The DNA was extracted by using extraction kit which gave relatively high yield concentration (40- 110 ng/µl), And purity within the optimal range. DNA concentration was adjusted to the proper concentration of PCR reaction. ABO gene was amplified by using two sets of specific primers for ABO locus with required alleles. Two separate segments of the glycosyltransferase gene containing nucleotide 261 in exon 6 and nucleotide 703 in exon 7 of ABO gene were amplified. The first set was used for the amplification of 468pb DNA fragment, Which contain the nucleotide 261. The second set was used to amplify 298bp DNA fragment, Which contain the nucleotide 703. Digestion products were run on 2% agarose gel, All fragments were compared with DNA ladder and there sizes were determined. The electrophoresis patterns of 100 samples showed that ABO genotypes were 5(4.0218%) AA, 23(23.9782%) AO, 2(3.3191%) BB, 24(22.6809%) BO, 8(8%) AB and 38(38%) OO. The O2 allele was encountered in this study. The study also showed that there is no difference when comparing the Phenotype with the Genotype.