L1 reaction mixture for molecular detection of Potato Leaf Roll Virus (PLRV) using quantitative RT-PCR

摘要

The present invention relates to a L1 reaction mixture for the molecular detection of potato coil virus (PLRV) by quantitative RT-PCR, which is characterized in that it contains 1 .mu.l of a sample of RNA tested, a reagent comprising a mixture of reverse transcriptase and DNA polymerase enzymes, a mixture of nucleotides, buffer containing magnesium chloride, MgCl.sub.2, sterile distilled water supplementing the reaction mixture to a final volume of 25 .mu.l primers with a specific nucleotide sequence of L-499F: AAG AAG TAT CCC TCA AAG CCT, L-675R: TTA GCG CGC CCT TGT AAA TCA AGC, 176 bp fragment, and L-567S probe: TG GAC GCT TGC GAG GTT GAT GAC TTT, the probe is labeled with fluorescent dye 6-carboxyfluorescein (FAM) and tetra is used as the quencher -methylcarboxyrhodamine (TAMRA).