Perbandingan Metode SYBR Green dan Hydrolysis Probe dalam Analisis DNA Gelatin Sapi dan Gelatin Babi Menggunakan Real Time Polymerase Chain Reaction

Zilhadia Zilhadia, Afifah Nurul Izzah, Ofa Suzanti Betha

Abstrak


Pemanfaatan gelatin secara luas menimbulkan kontroversi dan kekhawatiran bagi masyarakat muslim karena pada umumnya gelatin terbuat dari kulit babi dan sapi. Salah satu teknik analisis yang dapat membedakan gelatin sapi dan gelatin babi adalah Real Time Polymerase Chain Reaction (PCR). Real Time PCR merupakan metode analisis berbasis DNA yang handal, efektif, dan terpecaya. Dalam analisis kualitatif dan kuantitatif, Real Time PCR membutuhkan pewarna fluoresens. Pewarna fluoresens yang umum digunakan adalah SYBR green dan hydrolysis probe. Telah dilakukan perbandingan antara metode SYBR green dan hydrolysis probe dalam analisis DNA gelatin menggunakan Real Time PCR. DNA pada gelatin diisolasi menggunakan kit komersial. Isolat DNA gelatin sapi dan DNA gelatin babi didapatkan sebanyak 19,38 ng/μl dan 13,63 ng/μl dengan kemurnian 1,566 dan 1,573. Isolat DNA yang dianalisis dengan metode SYBR green menggunakan suhu annealing 65o C untuk primer sapi dan suhu annealing 60o C untuk primer babi. Isolat DNA yang dianalisis dengan metode hydrolysis probe menggunakan suhu annealing 60o C untuk primer babi dan primer sapi. Hasil analisis dari kedua metode menunjukkan bahwa metode hydrolysis probe lebih spesifik dalam mengidentifikasi DNA pada gelatin dibandingkan menggunakan metode SYBR green.


Kata Kunci


Gelatin; SYBR Green; Hydrolysis Probe; Real Time PCR

Teks Lengkap:

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Referensi


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DOI: https://doi.org/10.29208/jsfk.2017.4.1.194

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