Pembuatan DNA Ladder 100 bp Berbasis Polymerase Chain Reaction secara Mandiri

Abstract

Introduction. Polymerase chain reaction (PCR) is a rapid progress in molecular biology and is widely used in diagnosing diseases and genetic engineering. The PCR results interpretation needs an electrophoresis approach which requires a DNA ladder as an indicator of DNA size. Currently, Indonesia relies on commercial DNA ladders from abroad. Imported DNA ladder takes cost and transportation time. Therefore, Indonesia needs to produce DNA ladders. Objective. This study aims to produce homemade 100 bp DNA ladder. Methods. DNA from blood was isolated using the isolation kit (GIN-170, Sgima-Aldrich). Primer sets for 100, 200, 300, 400, 500, 600, 700, 800, 900, and 1000 bp were designed using Primer-BLAST. The expected DNA bands were produced by routine PCR and visualized using 2% (w/v) agarose gel electrophoresis. Amplicons were mixed to be a 100 bp DNA ladder. The homemade 100 bp DNA ladder was compared to the commercial one. Results and Discussion. The isolated DNA purity reached 1.878. Routine PCR produces bands of 100, 200, 300, 400, 500, 600, 700, 800, 900, and 1000 bp. The concentration of band 100, 500 and 1000 were 300 ng and the others were 200 ng. Mixing each band produces a 100 bp DNA ladder. Conclusion. A homemade 100 bp DNA ladder were produced using human blood DNA and has as good quality as a commercial one.