A 50-base pair DNA ladder from human DNA as a cost-effective molecular marker for health care researchers in low-resource laboratory settings.
Аннотация
INTRODUCTION
DNA ladders are predominantly purchased from manufacturers and tend to be more expensive than ladders created in laboratories. This study aimed to report on a practical method for preparing a 50-base pair (bp) DNA ladder using human DNA as a template.
METHOD: Human DNA from blood was isolated by spin column. Thirteen DNA fragments were amplified by polymerase chain reaction using designed primers and quantified. Amplicons were combined in 1.5-mL tubes. Laboratory-made and commercial 50-bp DNA ladders were compared qualitatively and quantitatively.
RESULTS
The isolated DNA purity was 1.89. The 50-bp DNA ladder obtained from human DNA exhibited good quality, with clearly visible fragments. Approximately 2 years after production, the laboratory-made DNA ladder's stability was acceptable, with a band intensity reduction of 5.45 arbitrary units compared with 5.58 arbitrary units observed in the commercial ladder. This protocol required only $20 in materials to produce a 50-bp DNA ladder for 100 runs, making it 4 to 15 times more economical than commercial products.
DISCUSSION: The laboratory-made DNA ladder is a useful standard in molecular research and has the potential to help health care professionals save both time and financial resources.
Предметные рубрики MeSH
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