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

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 tRNA leu gene mtDNA type 2 DM subject. 

Transfer RNA Mutation Associated with Type 2 Diabetes Mellitus

“…Various methods have been developed to detect the presence of the A3243G mutation or other mutations including high-performance liquid chromatography (HPLC), dot-blot hybridization, pyrosequencing, two-dimensional electrophoresis, peptide nucleic acid, electrochemical biosensors, radiolabeled polymerase chain reaction (PCR), ligation-mediated PCR (LM-PCR), allele-specific PCR, PCR amplification of specific alleles (PASA), PCR-restriction fragment length polymorphism (PCR-RFLP), and quantitative PCR (qPCR) [ 28 , 36 , 117 , 118 , 119 , 120 , 121 , 122 , 123 , 124 , 125 ]. Each of these methods has its advantages and limitations with some being easy to use but may be less sensitive, whereas others may involve the use of radioactive materials, which can be a concern.…”

In Silico Study of Mangostin Compounds and Its Derivatives as Inhibitors of α-Glucosidase Enzymes for Anti-Diabetic Studies

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