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2002, Cilt 32, Sayı 3-4, Sayfa(lar) 247-249
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The Importance of Touchdown-PCR in Detection of HGV-RNA
Ayhan Pekbay1, Meryem Çetin1, Abdülkerim Bedir2, Ahmet Saniç1, Murat Günaydın1
1Ondokuz Mayıs Üniversitesi Tıp Fakültesi, Mikrobiyoloji ve Klinik Mikrobiyoloji Anabilim Dalı, Samsun
2Ondokuz Mayıs Üniversitesi Tıp Fakültesi, Biyokimya Anabilim Dalı, Samsun
Keywords: Touchdown-PCR, optimization

Optimization is important in studies by using PCR. Touchdown (TD) PCR represents a fundamentally different approach to PCR optimisation. The optimisation is accomplished by focusing on a single variable, primer's annealing temperature. Moreover, a range of annealing temperatures is sequentially sampled during the course of a single cycling program.

In this study, hepatitis G virus RNA (HGV-RNA) was detected by using a RT-PCR with primers for 5'-non-coding region (5'-NCR) and (Non-structural-5a) NS5a genom region of HGV in 97 patients with chronic liver disease. We used TD-PCR technique to eliminate the non-spesific bands, because these had been seen in the PCR tests for primers of NS5a region whose melting temperatures were rather different from each other were used. Non-specific bands were quite decreased or disappeared, and bands belonging to desired products were clearly seen by the TDPCR cycle profile used.

The use of thermal cycler programming using of TD-PCR technique induces specificity and maximal productivity.


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