Optimization of DNA extraction of physic nut (Jatropha curcas) by selecting the appropriate leaf

EDI PRAYITNO, EINSTIVINA NURYANDANI

Abstract. Prayitno E, Nuryandani E. 2011. Optimization of DNA extraction of physic nut (Jatropha curcas) by selecting the appropriate leaf. Nusantara Bioscience 3: 1-6. Jatropha curcas L. has important roles as renewable source of bioenergy. The problem occurs on difficult of DNA extraction for its molecular breeding programs. The objectives of this research were to study which leaf best as source of DNA extraction. Four accession were used, namely J1 and J2 (Jawa Tengah), S1 (South Sumatra), and S2 (Bengkulu). First, third, fifth, seventh, and yellow leaves for each accession were extracted using modification of Doyle and Doyle (1987) method. Visualization and comparation with Lambda DNA, Spectrophotometer UV-Vis and cutting DNA with EcoRI enzyme were show quality and quantity of DNA. The result showed that third leaves have sufficient quality and quantity as source of DNA. Third leaves DNA quantity for J1 (19.33 µg/mL), J2 (26.21 µg/mL), S1 (31.20 µg/mL), dan S2 (61.03 µg/mL), and quality for each accession were 1.9063 (J1), 2.0162 (J2), 2.0116 (S1), and 2.0856 (S2).
Key words: Jatropha curcas, DNA extraction, appropriate, leaf.

Abstrak. Prayitno E, Nuryandani E. 2011. Optimalisasi ekstraksi DNA jarak pagar (Jatropha curcas) melalui pemilihan daun yang sesuai. Nusantara Bioscience. Nusantara Bioscience 3: 1-6. Jarak pagar (Jatropha curcas L.) mempunyai peran penting sebagai sumber bahan bakar nabati. Usaha pemuliaan tanaman ini secara molekuler sering terkendala sulitnya ekstraksi DNA. Penelitian ini bertujuan untuk mengetahui daun yang sesuai untuk digunakan sebagai sumber DNA. Penelitian ini dilakukan pada empat aksesi jarak pagar yaitu J1 dan J2 (Jawa Tengah), S1 (Sumatera Selatan), dan S2 (Bengkulu). Ekstraksi dilakukan pada daun pertama, ketiga, kelima, ketujuh, dan daun kuning dari setiap aksesi dengan metode Doyle and Doyle (1987) yang dimodifikasi. Kualitas dan kuantitas DNA hasil ekstraksi diketahui melalui visualisasi dengan pembanding DNA lambda, spektrofotometer UV-Vis pada panjang gelombang 260/280, dan pemotongan menggunakan enzim EcoRI. Hasil penelitian menunjukkan bahwa daun ketiga memadai untuk digunakan sebagai sumber DNA. Kuantitas DNA daun ketiga J1 (19,33 µg/mL), J2 (26,21 µg/mL), S1 (31,20 µg/mL), dan S2 (61,03 µg/mL). Sedangkan kemurniannya masing-masing yaitu 1,9063 (J1), 2,0162 (J2), 2,0116 (S1), dan 2,0856 (S2).
Kata kunci: Jatropha curcas, ekstraksi DNA, daun, sesuai.

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