1Chemistry Department, Faculty of Sciences and Mathematics, Diponegoro University, Indonesia
2Laboratorium Rekayasa Genetika, Pusat Antar Universitas Bioteknologi, Institut Teknologi Bandung, Indonesia
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@article{JKSA3321, author = {Mukhammad Asy’ari and A. Saifuddin Noer}, title = {Optimasi Konsentrasi MgCl2 dan Suhu Annealing pada Proses Amplifikasi Multifragmens mtDNA dengan Metoda PCR}, journal = {Jurnal Kimia Sains dan Aplikasi}, volume = {8}, number = {1}, year = {2005}, keywords = {PCR multifragments; mtDNA manusia Indonesia; optimasi PCR; MgCl2}, abstract = {Polymerase Chain Reaction (PCR) merupakan metode amplifikasi Deoxyribonucleic acid (DNA) secara invitro. Hingga saat ini para peneliti baru berhasil mengamplifikasi secara in vitro DNA mitokondria (mtDNA)manusia Indonesia sampai ukuran fragmen maksimal 1,6 kilobasa (kb) dan belum dilakukan untuk “banyakfragmen” (multifragments). Pada penelitian ini telah berhasil diamplifikasi multifragments DNA mitokondriamanusia Indonesia berukuran 5,4 kb; 4,2 kb; 3,5 kb; 2,1 kb; 1,6 kb dengan metode PCR standar melalui optimasikonsentrasi MgCl2 dan suhu annealing. Proses berlangsung dalam satu reaksi PCR menggunakan enzim taqpolimerase, lima primer yaitu dmt-2L, dmt-1H, FL, FH dan MH pada kondisi bufer standar dengan konsentrasiMgCl2 2,5 mM, selama 30 siklus pada suhu denaturasi 94oC (15 detik), annealing 65oC (30 detik) danextension 72oC (180 detik). Keberhasilan metode amplifikasi in vitro multifragments yang mengandung fragmenpanjang tersebut diharapkan bisa membuat proses amplifikasi beberapa gen dalam satu genom menjadi lebihcepat dan efisien.}, issn = {2597-9914}, pages = {23--27} doi = {10.14710/jksa.8.1.23-27}, url = {https://ejournal.undip.ac.id/index.php/ksa/article/view/3321} }
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