Perbandingan DNA Marker 50 bp Komersial dengan DNA Marker yang Bersumber dari DNA Manusia Menggunakan Fragmen DNA 50-1000 bp

Abstract

Introduction. The use of polymerase chain reaction (PCR) is a rapid improvement in the world of molecular biology. Visualization of PCR results using electrophoresis. Electrophoresis of PCR results requires DNA markers as an indicator of molecular weight measurement in units of base pairs (bp). Currently, Indonesia only relies on the production of commercial DNA markers from abroad. Commercial marker DNA has a relatively expensive price and a long delivery time due to long distances. Homemade marker DNA uses human DNA as the sample. Objective. This study aims to produce 50 bp DNA Marker using human blood samples. Methods. This research was conducted in July-September 2023 at the USU FK Integrated Laboratory. This research is a true experimental (pure experimental) with posttest only control group design. Primer design using NCBI Primer software. DNA from blood was isolated by spin column method. The concentration of DNA from PCR (amplicon) was measured with a nanophotometer. PCR specification with the determination of annealing temperature (Ta) was carried out by gradient PCR with 6 levels of temperature 55-60°C. Volume calculations were formulated specifically for each bp based on concentration. Amplicons of each bp are combined in a 1.5 mL tube with a certain volume into 50 bp Marker DNA. The quality of the homemade 50 bp Marker DNA was compared to qualitatively similar commercial 50 bp Marker DNA. Results and Discussion. The purity of the isolated DNA was 1.878. The optimal Ta at 50 bp and 100 bp was at 58oC, 150 bp-300 bp at 59oC, and 400-1000 bp at 60oC. PCR was again performed with the optimal Ta every bp. Each fragment produced one band. Conclusion. Homemade 50 bp DNA Marker already resembles commercial 50 bp DNA Marker.