Optimasi Konsentrasi MgCl2 dan Suhu Annealing pada Proses Amplifikasi Multifragmens mtDNA dengan Metoda PCR

Mukhammad Asy’ari; A. Saifuddin Noer

doi:10.14710/jksa.8.1.23-27

DOI: https://doi.org/10.14710/jksa.8.1.23-27

Optimasi Konsentrasi MgCl2 dan Suhu Annealing pada Proses Amplifikasi Multifragmens mtDNA dengan Metoda PCR

Mukhammad Asy’ari¹

, A. Saifuddin Noer²

¹Chemistry Department, Faculty of Sciences and Mathematics, Diponegoro University, Indonesia

²Laboratorium Rekayasa Genetika, Pusat Antar Universitas Bioteknologi, Institut Teknologi Bandung, Indonesia

Published: 1 Apr 2005.

Citation Format:

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.

Fulltext View|Download

Keywords: PCR multifragments; mtDNA manusia Indonesia; optimasi PCR; MgCl2

Article Metrics:

Article Info

Section: Research Articles

Language : ID

In Vol 8, No 1 (2005): Volume 8 Issue 1 Year 2005

Recent articles

Sintesis Bahan Hibrida Amino-Silika dari Abu Sekam Padi Melalui Proses Sol-Gel Pemucatan Minyak Sawit dengan Lempung Terpilar Sintesis 3-(3,4-Dimetoksi Fenil)-1-Propanol Melalui Hidroborasi Metileugenol Menggunakan H3B:Dietileter More recent articles

Most cited articles

Sintesis Metilisoeugenol dari Metileugenol Menggunakan Katalis KOH dalam Etanol Sintesis 3-(3,4-Dimetoksi Fenil)-1-Propanol Melalui Hidroborasi Metileugenol Menggunakan H3B:Dietileter Mekanisme Transport Lantanum Melalui Membran Cair Berpendukung (SLM) dengan Senyawa Pengemban campuran Tributyl Phosphate (TBP) dan Di(2-Etyl-Hexyl) Phosporic Acid (D2EHPA) Modifikasi Zeolit Alam dengan Ligan EDTA untuk Adsorpsi Ion Logam Pb2+ dan Cd2+ Asam Anakardat dari Kulit Biji Jambu Mete (Anacardium Occidentale L) yang Mempunyai Aktivitas Sitotoksik More cited articles

Barnes, M., Wayne. 1994, “PCR amplification of up to 35-kb DNA with high fidelity and high yield from  bacteriophage templates”, Pr°C. of the Natl. Acad. of Sci. of the USA 91: 2216-2220
Beck, S., 1998, “High Fidelity PCR: Enhancing the Accuracy of DNA Amplification”, The Scientist 12[1]: 17
Chamberlain, J.S., R.A. Gibbs, J.E. Ranier, P.N. Nguyen and C.T. Caskey., 1988, “Deletion screening of Duchenne muscular dystropy l°Cus via multiplex DNA amplification”, Nucleic Acids Res.16: 11141-11156
Cheng, S., and Kolmodin, L.A., 1997, XL PCR amplification of long targets from genomic DNA, Methods in Molecular Biology: PCR Cloning Prot°Cols, edited by Bruce A. White, 67, Humana Press, Totowa, New Jersey, page 17 – 29
Cheng, S., F°Ckler, C., Barnes, M.W., and Higuchi, R., 1994, “Effective amplification of long target from cloned inserts and human genomic DNA”, Pr°C. of the Natl. Acad. of Sci. of the USA 91: 5695-5699
Innis, A.M., and Gelfand, H.D., 1990, “Optimization of PCRs”, PCR Prot°Cols: A Guide to Methods and Applications
Newton, C.R., and Graham,A., 1997, PCR, 2nd edition, BIOS Scientific Publisher Limited, UK
Noer, A.S., Martasih, F., Mulyani, S., Muktiningsih, dan Wirahadi-kusumah, M., 1994, Analisis varian urutan nukleotida D-loop mtDNA manusia dari berbagai daerah di Indonesia, Proseeding Seminar Bersama UKM-ITB I, halaman 201-214
Puspasari, F., 1998, “Substitusi Enam Nukleotida Daerah Gen ND4, ND5 DNA Mitokondria Manusia Indonesia” , Tesis, Bidang Studi Magister Kimia Kimia Program Pascasarjana, ITB, Bandung
Ratnayani, K., 2000, Varian normal fragmen 1,6 kb daerah gen ND4 dan ND5 DNA mitokondria 10 individu Indonesia, Tesis, Bidang Khusus Biokimia, Program Studi Kimia, Program Pascasarjana, ITB, Bandung

Last update:

Optimization of the Detection Method for Xanthomonas axonopodis on Chili Pepper Seeds Using Polymerase Chain Reaction with Two Difference Primers
D Aisya, M E Poerwanto, D Wicaksono, J D Ortuoste, A. Loch, R.C. Joshi, A. Munawar, M.T. Multazam. BIO Web of Conferences, 69 , 2023. doi: 10.1051/bioconf/20236901033
Proceedings of the 3rd International Conference on Biology, Science and Education (IcoBioSE 2021)
Zulyusri Zulyusri, Rosi Fitri Ramadhani, Desyanti Desyanti. Advances in Biological Sciences Research, 32 , 2023. doi: 10.2991/978-94-6463-166-1_22

Last update: 2024-11-12 01:09:34

No citation recorded.

As an article writer, the author has the right to use their articles for various purposes, including use by institutions that employ authors or institutions that provide funding for research. Author rights are granted without special permission.

Author who publishes a paper at JKSA has the broad right to use their work for teaching and scientific purposes without the need to ask permission, including: used for (i) teaching in the author's class or institution, (ii) presentation at meetings or conferences and distributing copies to participants ; (iii) training conducted by the author or author's institution; (iv) distribution to colleagues for research use; (v) use in the compilation of subsequent authors' works; (vi) inclusion in a thesis or dissertation; (vi) reuse of part of the article in another work (with citation); (vii) preparation of derivative works (with citation); (viii) voluntary posting on open websites operated by authors or author institutions for scientific purposes (follow the CC BY-SA License).

Authors and readers can copy and redistribute material in any media or format, and mix, modify, and build material for any purpose but they must provide appropriate credit (provide article citation or content), providing links to the license, and indicate if there are changes.

The authors submitting a manuscript do so on the understanding that if accepted for publication, copyright of the article shall be assigned to Jurnal Kimia Sains dan Aplikasi (JKSA). Copyright encompasses rights to reproduce and deliver the article in all form and media, including reprints, photographs, microfilms and any other similar reproductions, as well as translations.

Reproduce any part of this journal, its storage in the database or its transmission by all forms or media is permitted does not need for written permission from JKSA. However, it should be cited as an honor in academic manners

JKSA and the Chemistry Department of Diponegoro University and the Editor make every effort to ensure that there are no data, opinions, or false or misleading statements published in JKSA. However, the content of the article is the sole and exclusive responsibility of each author.

The Copyright Transfer Form can be downloaded here: [Copyright Transfer Form - Indonesian] [Copyright Transfer Form - English]. The copyright form should be signed originally and send to the Editor in the form of printed letters, scanned documents sent via email or fax.

Adi Darmawan, Ph.D (Editor in Chief)

Editor in chief of Jurnal Kimia Sains dan Aplikasi (JKSA)

Chemistry Department, Faculty of Sciences and Mathematics, Diponegoro University

Jl. Prof. Soedarto, Kampus Undip Tembalang, Semarang, Central Java, Indonesia 50275
Email: jksa@live.undip.ac.id