PCR – RFLP amplification applied to the determination of ANXA2 gene single nucleotide polymorphism in sickle cell patient.

Abstract

PCR-RFLP is a generally applicable technique for the detection of known single nucleotide polymorphism. Here we are presenting to the application of the PCR-RFLP technique to determination of the ANXA2 gene single nucleotide polymorphism and their frequency among Asian Indian sickle cell patients. Five known SNPs; rs7170178, rs73435133, rs73418020, rs72746635 and rs73418025 of ANXA2 gene determined using the HpyCH4V, DdeI, HpyCH4III, and Sau 961 restriction enzyme. Restriction enzyme DdeI used for both SNPs rs73435133 and rs72746635. However all individual among patient and control groups except rs7170178; were wild type and frequency was zero. ANXA2 SNP- rs7170178; heterozygous frequency increased among patient groups especially sickle homozygous patients. However the p-value was not significant (0.41 and 0.93) for heterozygous and homozygous respectively. Statistical analysis was performed on graph-pad statistics software. Yates’ chi-square test was used to assess inter-group significance.

Keywords

• SNPs
• PCR-RFLP Restriction digestion
• ANXA2