Reaction mixture for molecular detection of potato spindle tuber viroid disease in potato tubers by employing quantitative RT-PCR

摘要

The present invention relates to a reaction mixture for molecular detection of potato spindle tuber viroid disease in potato tubers by use of quantitative RT-PCR, which reaction mixture is characterized in that it contains 1 ml of a RNA sample being tested, 12.5 ml of 2x master mix containing a buffer mixture, DNA polymerase, deoxyribonucleotides, and magnesium chloride, 1.0 ml of a mixture of reverse transcriptase and RNAse inhibitor, further 9.3 ml of a sterile distilled water completing the reaction mixture to a final volume of 25 ml and 0.1 ml of each primer with concentration of 100miM and a specific nucleotide sequence QRT R_PSTVd: CTT TCT TCg ggT gTC CTT CC, QPCR R_PSTVd: CTC ggg AgC TTC AgT TgT TTC, which amplify a fragment with the size of 101 bp, and 1 ml of PSTVd probe with concentration of 30 mM and a specific nucleotide sequence TgCgCTgTCgCTTCggCTACT, wherein said probe is labeled with a fluorescence dye 6-carboxyfluoresceine (FAM) and tetramethylcarboxyrhodamine (TAMRA)i is used as an extinguisher.