Total RNA extraction method and Prunus species influence the detection of Plum pox potyvirus by real-time RT-PCR | Rastlinska vrsta in metoda izolacije celokupne RNA vplivata na uspesnost detekcije virusa sarke s PCR v realnem casu
2011
Mavric-Plesko, I., Agricultural Inst. of Slovenia, Ljubljana (Slovenia) | Virscek-Marn, M., Agricultural Inst. of Slovenia, Ljubljana (Slovenia) | Toplak, N., Agricultural Inst. of Slovenia, Ljubljana (Slovenia)
Successful total RNA (totRNA) extraction is a prerequisite for a successful real-time PCR. In the present work we compared one manual and one automated totRNA extraction method for detection of Plum pox potyvirus (PPV) in leaves of different Prunus sp. using developed real-time RT-PCR assay. Advantages and disadvantages of compared methods are described in the view of sensitivity, reproducibility and in terms of laboratory use. The results suggest that the column based totRNA extraction method is more effective for apricot, plum and peach. In the case of damson the differences in real-time PCR results between both extraction methods were negligible. In case of negative results obtained with automated method, manual column based extraction method should be used additionally.
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