Optimization of cDNA amplification of Apricot Latent Virus (ApLV) from various plant tissues sources.
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.
Gumus
M
M
Department of Plant Protection, Faculty of Agriculture, Ege University, 35100 Bornova, Izmir, Turkey.
Sipahioğlu
H M
HM
Paylan
I C
IC
Erkan
S
S
D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics*; D052677:Flexiviridae / Q000235:genetics*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods; D010944:Plants / Q000821:virology*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
D017931:DNA Primers; D018076:DNA, Complementary