KOMPONEN, TAHAPAN DAN VARIASI POLIMERASE CHAIN REACTION : ARTIKEL REVIEW
DOI:
https://doi.org/10.31004/jkt.v6i1.43342Keywords:
PCR, DNA polimerase, amplifikasi DNA, Real-Time PCR, RT-PCRAbstract
Polymerase Chain Reaction (PCR) merupakan teknik perbanyakan DNA secara in vitro yang memanfaatkan enzim DNA polymerase termostabil. PCR memiliki peran penting dalam berbagai bidang, terutama biomedis dan forensik, karena kemampuannya dalam mendeteksi serta mengidentifikasi DNA target dengan sensitivitas dan spesifisitas tinggi. Penelitian ini bertujuan untuk menjelaskan prinsip kerja PCR, komponen utama dalam reaksi PCR, tahapan PCR, serta variasi teknik PCR yang berkembang, seperti Real-Time PCR, Reverse Transcription-PCR (RT-PCR), dan direct PCR. Metode yang digunakan adalah studi literatur terhadap berbagai sumber ilmiah relevan. Hasil penelitian menunjukkan bahwa PCR terdiri dari tiga tahap utama, yaitu denaturasi, annealing, dan elongasi, yang dilakukan secara berulang-ulang hingga menghasilkan jumlah salinan DNA yang cukup untuk dianalisis. Berbagai variasi PCR telah dikembangkan untuk meningkatkan sensitivitas dan efisiensi deteksi. Real-Time PCR memungkinkan kuantifikasi DNA secara akurat, RT-PCR memungkinkan analisis RNA melalui konversi menjadi DNA, sementara direct PCR menyederhanakan proses dengan menghilangkan tahap ekstraksi DNA. Aplikasi PCR sangat luas, termasuk dalam diagnostik penyakit infeksi, forensik, penelitian genetik, deteksi mutasi genetik, identifikasi mikroorganisme, dan pengujian makanan. Keunggulan PCR dalam kecepatan, ketepatan, serta kemampuannya mendeteksi DNA dalam jumlah sangat kecil menjadikannya alat yang sangat penting dalam ilmu biologi molekuler. Kesimpulannya, PCR merupakan teknik fundamental dalam biologi molekuler yang terus berkembang, memberikan kontribusi besar dalam penelitian dan aplikasi klinis.References
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