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All Journal SCRIPTA SCORE Scientific Medical Journal eJournal Kedokteran Indonesia Makara Journal of Science
Nasution, Luthfi Umam Hakim
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Biochemistry, Genetics & Molecular Biology Immunology & microbiology Medicine & Pharmacology Neuroscience Public Health

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Journal : Makara Journal of Science

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Lab-made 100 bp DNA Ladder using Polymerase Chain Reaction and Human DNA Bariqi, Muhammad Ilmam; Yamamoto, Zulham; Firjatu, Putri Chalya; Nasution, Luthfi Umam Hakim; Lubis, Oryza Sativa
Makara Journal of Science Vol. 30, No. 1
Publisher : UI Scholars Hub

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Abstract

Polymerase chain reaction (PCR) is a rapid, molecular biology technique widely used in disease diagnosis and genetic engineering. Conventional PCR products require agarose gel electrophoresis, which employs a DNA ladder as a size reference. Most commercial ladders are plasmid-based and reliable but require additional culture time. We suggest a more efficient method for producing a DNA ladder using DNA derived from human blood. DNA was isolated using a commercial kit. Primer sets generating 100–1000 base pair (bp)-long fragments bearing target regions p12, p13, and p14 were designed using Primer-BLAST. DNA was amplified by routine PCR, visualized on a 1% (w/v) agarose gel, and pooled to form a 100-bp ladder. The lab-made ladder was compared with a commercial ladder. Extracted DNA showed a purity of 1.878. Bands of 100, 500, and 1000 bp were added in greater amounts to produce wider, more visible bands. The pooled fragments served as a functional 100-bp DNA ladder. A key limitation is the lack of sequencing, as genomic variations may affect flanking regions and potentially alter amplicon sizes. A 100-bp DNA ladder generated using DNA extracted from human blood had good quality, comparable to that of a commercial ladder.
Co-Authors Bariqi, Muhammad Ilmam Darlan, Dewi Masyithah Firjatu, Putri Chalya Lubis, Oryza Sativa Pane, Baharudin Yusuf Panggabean, Grestia Angraini Perdana, Muhammad Agung Pratama, Alfath Bagus Samosir, Fauzan Azmi Hasti Habibi Simanjuntak, Samuel Manutur Siregar, Moammer Saddam Syahputra Yamamoto, Zulham