METODE EKSTRAKSI DNA UNTUK DETEKSI MOLEKULER

Rosy Hutami, N Idzni, R Ranasasmita, Mira Suprayatmi

Abstract


Dalam teknik deteksi molekuler seperti Loop-Amplification Mediated Polymorphism (LAMP) dan Polymerase Chain Reaction (PCR), pembuatan hulu asam deoksiribonukleat (DNA) sangat penting. Ekstraksi fase cair dan ekstraksi fasa padat merupakan beberapa metode ekstraksi DNA yang tersedia. Tujuan dari penelitian ini adalah untuk mengkarakterisasi metode dan produk ekstraksi DNA berdasarkan kemurnian DNA, visualisasi DNA, konsentrasi DNA, dan waktu pemrosesan metode ekstraksi DNA. Metode ekstraksi yang dievaluasi meliputi metode fenol-kloroform (Metode A) sebagai ekstraksi fasa cair dan metode Ekstraksi Surefood kit (Metode B) sebagai ekstraksi fasa padat. Hasil penelitian menunjukkan bahwa Metode A dapat dilakukan pada sampel dengan konsentrasi DNA sangat rendah berkisar antara 7,00 sampai 9,45 ng / μl dengan kemurnian yang baik (1,80-2,10). Meski tidak menunjukkan DNA isolat band pada gel agarosa 1% dan membutuhkan waktu pemrosesan ± 30 jam. Metode B memiliki performa yang baik dalam mengekstraksi sampel dengan DNA dengan konsentrasi tinggi (49,67 sampai 357,28 ng / μl) dengan kemurnian yang baik (1,93 sampai 2,07). Metode ini menunjukkan band untuk setiap sampel DNA pada gel agarose 1% dan membutuhkan waktu ±1 jam. Kedua metode tersebut dapat digunakan untuk preparasi sampel dalam analisis molekuler termasuk tujuan otentikasi halal.

KATA KUNCI: fenol, kloroform, Surefood kit, LAMP

 

 

DNA EXTRACTION METHOD FOR MOLECULAR DETECTION


ABSTRACT

In molecular detection technique such as Loop-Amplification Mediated Polymorphism (LAMP) and Polymerase Chain Reaction (PCR), right upstream preparation of deoxyribonucleic acid (DNA) is very important. Liquid phase extraction and solid phase extraction are some of DNA extraction methods those are available. The purpose of this research was to characterizedthe method and product of DNA extraction based on DNA purity, DNA visualization, DNA concentration, and processing time of DNA extraction methods. Extraction methods evaluated included phenol-chloroform method (Method A)as liquid phase extraction and Surefood Extraction kit method (Method B) as solid phase extraction. Result showed that Method A could be performed on samples with very low DNA concentrations ranging from 7.00 to 9.45 ng/µl with a good purity (1.80 to 2.10). Although,  it showed no DNA isolates bands on gel agarose 1% and need ± 30 hours processing time. Method B had a good performa in extracting sample with high concentration DNA (49.67 to 357.28 ng/µl) with a good purity (1.93 to 2.07). This method showed bands for each DNA samples on gel agarose 1% and need about ± 1 hour processing time. Both methods can be used for sample preparation in molecular analysis including halal authentication purposes.

 

 


Keywords


phenol, chloroform, Surefood kit, LAMP.

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DOI: https://doi.org/10.30997/jp.v8i2.1056

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