Optimization of cDNA Amplification of Apricot Latent Virus (ApLV) from Various Plant Tissues Sources
Makaleler > Tam Makale · 2007
Yazarlar: M. Gumus, H.M. Sipahioglu, İsmail Can PAYLAN, S. Erkan
Pakistan Journal of Biological Sciences, Uluslararası, Scopus, 2007, cilt 10, ss 936-940
DOI: 10.3923/pjbs.2007.936.940
Özet
Although the reverse transcriptase polymerase chain reaction (RT-PCR) procedure is basically simple operation, often it is not possible to achieve optimum results without optimizing the protocols. An RT-PCR method targeting a 200 bp sequence of the CP gene of Apricot Latent Virus (ApLV) was used as a model to improve the detection limit and to compare the behavior of three different plant tissues in a RT-PCR assay. A number of factors should be considered when selecting the optimal system for RT-PCR. Important considerations include the optimal concentrations of MgCl2, dNTP, Taq DNA polymerase enzyme, specific primer and the amount of cDNA for the downstream applications. This study therefore discusses a series of critical PCR parameters and feasible strategies for optimization of RT-PCR detection of ApLV.