Desain primer untuk identifikasi daging katak menggunakan teknik pentaplex PCR
Keywords:
halal product assuranc, jaminan produk halal, in sillico, frog, katak, PCR primer, primer PCRAbstract
Frog is non-halal food source that is vulnerable to be used as halal food replacement or food adulteration. Therefore, this study’s purpose was to design the primer for identifying the presence of frog meat in food product using pentaplex PCR technique. The primers were designed based on the sequence of cytochrome b gene from frog (Rana temporaria) stored in gene bank NCBI. PrimerBlast was used as the in sillico tool for primer designing using some parameters such as aplicons length and Tm value of the primer. Ten primer pairs were produced as candidates to be used in the PCR analysis, but the chosen five pairs, namely primer 1, 2, 4, 7, and 10, were based on the amplification region from each primer pair. The region from those 5 primers were expected to boost the sensitivity of pentaplex PCR as the development method for halal food assurance.
Katak merupakan salah satu bahan pangan non-halal yang rentan digunakan untuk pemalsuan atau campuran bahan pangan. Maka dari itu, ujuan dari penelitian ini adalah untuk mendesain primer yang dapat digunakan untuk mengidentifikasi keberadaan daging katak dalam produk olahan pangan menggunakan teknik pentaplex PCR. Primer didesain berdasarkan urutan gen sitokrom b dari katak (Rana temporaria) yang dapat diakses pada gen bank NCBI. Desain primer tersbeut dilakukan secara in sillico menggunakan PrimerBlast dengan beberapa parameter seperti panjang amplikon dan nilai Tm dari primer. Dari 10 kandidat primer yang diperoleh, dipilih 5 primer yang akan digunakan sebagai primer untuk pentaplex PCR yakni pasangan primer nomor 1, 2, 4, 7, dan 10 yang didasari pada daerah amplifikasi dari masing-masing pasangan primer. Kelima primer tersbeut diharapkan tidak hanya mencakup hampir keseluruhan gen sitokrom b katak tetapi juga meningkatkan sensitivitas sebagai upaya penjaminan produk halal.
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