Molecular Investigation of Genetic Polymorphisms in Type 2 Diabetic Patients Using Random Amplified Polymorphic DNA (RAPD-PCR)

Abstract

To find DNA markers associated with Type 2 diabetes mellitus (T2D) using RAPD-PCR (Random amplified polymorphic DNA-Polymerase Chain Reaction). Peripheral blood samples were collected from 12 unrelated Iraqis with type 2 diabetes mellitus and 10 apparently healthy individuals (control subjects). DNA was extracted and amplified using RAPD-PCR. Out of the 16 primers used, 3 did not produce amplification patterns. Seven primers produced monomorphic bands while 6 primers namely A10, A18, C5, D20, R3 and R4 produced polymorphic DNA profiles. The highest discriminatory power was produced by primers A18 and D20 reaching 25%. Primer D20 produced the highest number of bands (16) and largest molecular weight band 2.470Kb whilst primer A10 produced the lowest number of bands (3) and primer R4 the smallest molecular weight band of 0.296Kb. Furthermore a band of 1.056 Kb was produced by primer R4 with frequency of 100% in diabetic patients and total absence in control subjects. The total length of the genome screened was approximately 118.661Kb, 40% of which represent polymorphisms. In conclusion Polymorphisms between diabetic patients and control subjects can be detected by RAPD-PCR and DNA markers associated with type 2 diabetes mellitus can be found using the same technique