KONSENTRASI DAN KEMURNIAN DNA HASIL EKSTRAKSI RUMPUT BENGGALA
DNA CONCENTRATION AND PURITY FROM BENGGALA GRASS EXTRACTION
Abstract
Rumput benggala (Panicum maximum) merupakan hijauan pakan unggul di wilayah tropis yang memerlukan analisis molekuler untuk mendukung program pemuliaan dan identifikasi genetik. Tahap awal yang krusial dalam analisis tersebut adalah ekstraksi DNA genom berkualitas tinggi yang bebas dari kontaminan. Penelitian ini bertujuan untuk mengevaluasi efektivitas metode ekstraksi DNA menggunakan DNA Mini Kit for Plant GP 100 (Geneaid) dengan modifikasi prosedur pada tiga kultivar rumput benggala (Panicum maximum cv. Purple Guinea, cv. Common, dan cv. Gatton). Modifikasi metode yang diterapkan meliputi stabilisasi sampel melalui penyimpanan pada suhu dingin selama 24 jam sebelum penggerusan dan optimasi pengeringan GD column melalui sentrifugasi pada kecepatan tinggi (13.000 rpm selama 6 menit). Peubah yang diamati adalah konsentrasi dan kemurnian DNA menggunakan spektrofotometer NanoDrop pada panjang gelombang 260 nm dan 280 nm. Hasil penelitian menunjukkan bahwa terdapat variasi efisiensi ekstraksi antarkultivar. Kultivar Purple Guinea menghasilkan rata-rata konsentrasi DNA tertinggi sebesar 92,15 ng/µL dengan indeks kemurnian (A260/A280) 1,87. Kultivar Gatton menghasilkan konsentrasi sebesar 83,78 ng/µL dengan nilai kemurnian tertinggi dan paling konsisten, yaitu 1,90. Sementara itu, kultivar Common menghasilkan konsentrasi terendah sebesar 42,10 ng/µL dengan rata-rata kemurnian 1,82, serta menunjukkan variabilitas kualitas antarsampel yang lebih lebar. Modifikasi metode ekstraksi menggunakan Mini Kit terbukti efektif menghasilkan DNA berkualitas tinggi yang memenuhi standar untuk aplikasi molekuler lanjutan, di mana kultivar Purple Guinea dan Gatton menunjukkan hasil purifikasi terbaik.
ABSTRACT
Benggala or Guinea grass (Panicum maximum) is a superior forage crop in tropical regions that requires molecular analysis to support breeding and genetic identification programs. A crucial initial step in this analysis is the extraction of high-quality genomic DNA free of contaminants. This study aimed to evaluate the effectiveness of a DNA extraction method using the DNA Mini Kit for Plant GP 100 (Geneaid) with procedural modifications on three guinea grass cultivars (Panicum maximum cv. Purple Guinea, cv. Common, and cv. Gatton). The modifications applied included sample stabilization through cold storage for 24 hours prior to grinding and optimization of GD column drying via high-speed centrifugation (13,000 rpm for 6 minutes). The parameters observed were DNA concentration and purity using a NanoDrop spectrophotometer at wavelengths of 260 nm and 280 nm. The results showed variations in extraction efficiency among cultivars. The Purple Guinea cultivar yielded the highest average DNA concentration at 92.15 ng/µL with a purity index (A260/A280) of 1.87. The Gatton cultivar produced a concentration of 83.78 ng/µL with the highest and most consistent purity value of 1.90. Meanwhile, the Common cultivar produced the lowest concentration at 42.10 ng/µL with an average purity of 1.82, displaying wider quality variability among samples. The modified mini kit extraction method proved effective in generating high-quality DNA that meets the standards for advanced molecular applications, with Purple Guinea and Gatton cultivars demonstrating the best purification results.
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