Polimeraz zincir reaksiyonu

DNA'nın genetik bilgiyi taşıdığının ortaya konması, canlıyı araştıran yöntemlerin moleküler düzeye yönelmesine neden olmuştur. Moleküler genetik teknikler sayesinde birçok hastalığın kesin ve hızlı teşhisi yapılmakta ve genetik tedavi yöntemleri araştırılmaktadır. Fötal dönem hastalıkları, DNA parmak izi çalışmaları ve ailesel kanser taramaları mümkün olmaktadır. Babalık tayini, cinayet vakalarının aydınlatılması gibi adli tıp alanında da genetik analizler yapılmaktadır. DNA'nın hızlı ve güvenilir bir şekilde çalışılmasına imkan veren polimeraz zincir reaksiyonunun (PCR) keşfi ve takip eden yıllarda geliştirilmesi ile moleküler genetik alanındaki gelişmeler ivme kazanmıştır. Bu derlemede; PCR tekniğinin temel prensibi, işleyişi ve kullanılabileceği sahalar hakkında bilgi sunulmaktır.

Anahtar Kelimeler:

DNA parmakizi, tanı teknikleri, moleküler genetik, polimeraz zincir reaksiyonu, teknikler, kullanım yerleri

Polymerase chain reaction

The discovery of DNA's genetic information function led the methods that study living beings to move to the molecular level. By means of molecular genetic technics many of diseases have been diagnosed definitely and genetic therapy methods have been improved. The health of fetus can be determined before the birth and by the DNA fingerprinting studies familial cancer researchs can be made. Genetic studies such as paternity determination and enlightening crime events in forensic medicine can be made. The investigation and development of polymerase chain reaction (PCR) allow DNA studies to be safe and rapid. The aim of this review is to give information about basic principles of process and usage of PCR technics.

Keywords:

DNA fingerprinting, diagnostic techniques, molecular genetics, polymerase chain reaction, techniques, uses,

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1.Lehninger AL: Biochemistry. 309-331, Kalyani Publishers, New Delhi-1995.
2.Kaya N: Biyokimya.208-219, Atatürk Üniversitesi Yayınlan No:743, Erzurum-1993.
3.Temizkan G, Arda N: Moleküler Biyolojide Kullanılan Yöntemler. 57-66, BİYOGEM Yayınları No:l, Nobel lıp Kitabevleri, İstanbul-1999.
4.Alberts B, Bray D, Lewis J, Raff M, Roberts K, ™son JD: Molecular Biology of the Cell. 3rd Edition, 316-31 /, Garland Publishing, New York-1994.
5.Şahin F, Çiftçi M, Pirim İ: Polimeraz zincir reaksiyonu (PCR). II. Uygulamalı Moleküler Biyoloji Teknikleri Kurs Notlan. AÜ Biyoteknoloji Uygulama ve Araştırma Merkezi, 2000.
6.Dieffenbach CW, Dveksler GS: PCR Primer: A Laboratory Manual. 1-287, Cold Spring Harbor Laboratory Press,19.ew York-1995.
7.Taylor GR, Robinson P: The polymerase chain reaction. From Functional Genomics to High-School Practical Classes. Current Opinion in Biotechnology, 9: 35-42, 199o.
8.Persing DH: Polymerase chain reaction: Trenches to Benches. J Clin Microbiol, 29:7,1281-1285, 1991.
9.Erlich HA, Gelfand D, Sninsky JJ: Recent advances in the polymerase chain reaction. Science, 252: 1643-1651, 1991.
10.Taylor GR, Logan WP: The polymerase chain reaction: New variations on an old theme. Current Opinion in Biotechnology, 6: 24-29, 1995.
11.Taga Y, Aslan D, Güner G, Kutay FZ: Tıbbi Laboratuvarlarda Standardizasyon ve Kalite Yönetimi. 426- 434, Mart Matbaacılık A.Ş., 2001.
12.Howe CJ, Ward ES: Nucleic acids sequencing: A practical approach. 53-114, IRL Press at Oxford University Press, Oxford-1989.
13.Innis MA, Gelfand DH, Sninsky JJ: PCR Strategies. 39-67, Academic Press, San Diego-1995.
14.McPherson MJ, Hames BD, Taylor GR: PCR 2: A Practical Approach. 7-118, Oxford University Press, Oxford-1995.
15.Banks M: DNA restriction fragment length polymorphism among British isolates of Aujeszky's Disease virus: Use of the polymerase chain reaction to discriminate among strains. Br VetJ, 149, 155-162, 1993.
16.Johnson JR: Development of polymerase chain reaction- based assays for bacterial gene detection. J Mic Meth, 41, 201- 209, 2000.
17.Mandrell RE, Wachtel MR: Novel detection techniques for human pathogens that contaminate Poultry. Curr Opi Biot 10, 273-278, 1999.
18.Grainger J, Madden D: The polymerase chain reaction: turning needles into haystacks. Biologist, 40:5, 197-200,1993.
19.Tag CG, Gressner AM, Weiskirchen R: An unusual melting curve profile in LightCycler multiplex genotyping of the hemochromatosis H63D/C282Y gene mutations. Clin Bioc 34, 511-515,2001.
20.Dixon AK, Richardson PJ, Pinnock RD, Lee K: Gene- Expression Analysis at the Single-Cell Level. TIPS, 21: 65-70, 2000.
21.Schalkwyk LC, Francis F, Lehrach H: Techniques in mammalian genome mapping. Current Opinion in Biotechnology, 6: 37-43, 1995.
22.Orlando V: Mapping Chromosomal Proteins in vivo by Formaldehyde-Crosslinked-Chromatin Immunoprecipitation. TIPS, 25: 99-103, 2000.
23.Press RD, Deloughery TG: Trombotik risk değerlendirmesi: Sık rastlanan, tedavi edilebilir genetik bir hastalık için moleküler ve konvansiyonel testler. Biyokimya Dergisi, 3: 42- 46,1999.
24.Bustin S, Dorudi S: Molecular Assesment of Tumour Stage and Disease Recurrence Using PCR-based Assays. Molecular Medicine Today, 389-396,1998.
25.Pommepuy M, Le Guyader F: Molecular approaches to measuring microbial marine pollution. Current Opinion in Biotechnology, 9:292-299, 1998.
26.Jansson JK: Tracking genetically engineered microorganisms in nature. Current Opinion in Biotechnology, 6: 275-283, 1995.
27.Rashtchian A: Novel methods for cloning and engineering genes using the polymerase chain reaction. Current Opinion in Biotechnology, 6: 30-36, 1995.
28.Davis LG, Dibner MD, Battey JF: Basic Methods in Molecular Biology. 23-58, Elsevier, New York-1986.
29.Lie YS, Petropoulos CJ: Advances in Quantitative PCR Technology: 5' Nuclease Assays. Current Opinion in Biotechnology, 9: 43-48, 1998.
30.Sterky F, Lundeberg J: Sequence analysis of genes and genomes. Journal of Biotechnology, 76: 1-31, 2000.
31.Zhou H, Liu W, Lamont SJ: Genetic variation among chicken lines and mammalian species in spesific genes. Poultry Science, 80, 284-288, 2001.
32.Tillib SV, Mirzabekov AD: Advances in the analysis of DNA sequence variations using oligonucleotide microchip technology. Curr Opi Biot, 12, 53-58, 2001.
33.Newton CR: PCR Essential Data. 1-56, John Wiley & Sons Ltd, Chichester-1995.