Desain Primer In Silico Untuk Analisis Expresi Gen Osteopontin (OPN) Sebagai Penanda Kesuburan Pada Kambing Jantan (Capra hircus)
Abstrak
Tujuan dari penelitian ini adalah untuk mendesain sepasang primer agar dapat digunakan dalam analisis ekspresi gen OPN sebagai penanda kesuburan pada kambing jantan (Capra hircus). Metode penelitian yang digunakan adalah kajian molekuler secara in silico berbasis bioinformatika. Data nukleotida diambil dari situs NCBI dengan kode NM_001285667.1. Data yang diperoleh kemudian diolah dengan menggunakan Primer-Blast pada situs NCBI untuk memperoleh kandidat primer. Kandidat primer Selanjutnya dianalisis struktur sekunder dan dimer primer menggunakan OligoEvaluator Sigma-Aldrich. Primer yang dirancang kemudian dianalisis spesifisitas primer dan tingkat kesamaan sekuens dengan database nukleotida yang ada di NCBI menggunakan Primer-Blast dan Nucleotide-Blast pada situs NCBI. Hasil penelitian menunjukkan bahwa diperoleh satu pasangan primer yang memiliki kriteria ideal primer yaitu primer forward 5’- ACCCTCCCGAGTAAGTCCAA-3’ dan reverse 5’- AGTCCTCCTCTGTGGCATCT-3’ yang mampu mengamplifikasi daerah gen OPN dengan panjang produk 325 bp. Hasil uji kinerja secara in silico juga menunjukkan bahwa primer tersebut dapat digunakan untuk deteksi gen OPN Capra hircus secara spesifik.
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