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Restriction Enzymes ApaI Analysis to Find A3243G Mutation in Indonesia Diabetes Mellitus Type II Patients
Richie Agusta Iwan Chandra, Sriwidodo, Ajeng Diantini, Iman P. Maksum
Universitas Padjadjaran, Journal of Medical and Bioengineering Vol. 4, No. 6, December 2015
Bahasa Inggris
Universitas Padjadjaran, Journal of Medical and Bioengineering Vol. 4, No. 6, December 2015
DNA mutation, MIDD, mtDNA A3243G, PCR-RFLP, restriction enzyme ApaI
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 tRNA leu gene mtDNA type 2 DM subject.