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Restriction Enzymes ApaI Analysis to Find A3243G Mutation in Indonesia Diabetes Mellitus Type II Patients

Richie Agusta Iwan Chandra1, Sriwidodo1, Ajeng Diantini1 , and Iman P. Maksum2
1.Pharmacy Faculty Universitas Padjadjaran, Sumedang, Indonesia
2.Chemistry Departement Mathematics and Natural Science Faculty Universitas Padjadjaran, Sumedang, Indonesia
Abstract—The Use of PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism) to find out the potential of mitochondrial DNA mutation at A3243G in type II Diabetic Patient has been done. Peripheral blood from 100 Indonesian type 2 diabetic subjects was selected randomly for this experiment. Peripheral blood lymphocyte was isolated, lysed, and it was in vitro amplified by PCR using a pair D1/D2 primers. PCR products were 294 base pair (bp) fragments which were then purified by ethanol precipitation method and characterized by restriction enzyme ApaI. Heteroplasmic A3243G mutation which was identified in 2 Subject (0,02%) was shown by 3 electrophoretic bands, 2 restriction products of APAI, i.e a 182 bp fragment and a 112 bp fragment; also a full fragment 294 bp, this means show that PCR-RFLP technique was approved for identifying heteroplasmic A3243g mutation in a tRNAleu gene mtDNA type 2 DM subject.

Index Terms—DNA mutation, mtDNA A3243G, MIDD, restriction enzyme ApaI, PCR-RFLP

Cite: Richie Agusta Iwan Chandra, Sriwidodo, Ajeng Diantini, and Iman P. Maksum, "Restriction Enzymes ApaI Analysis to Find A3243G Mutation in Indonesia Diabetes Mellitus Type II Patients," Journal of Medical and Bioengineering, Vol. 4, No. 6, pp. 492-496, December 2015. Doi: 10.12720/jomb.4.6.492-496
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