Optimization of cDNA Amplification of Apricot Latent Virus (ApLV) from Various Plant Tissues Sources
Articles > Article · 2007
Authors: M. Gumus, H.M. Sipahioglu, İsmail Can PAYLAN, S. Erkan
Pakistan Journal of Biological Sciences, International, Scopus, 2007, vol 10, pp 936-940
DOI: 10.3923/pjbs.2007.936.940
Summary
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.