Molecular assays to detect the presence and viability of Phytophthora ramorum and Grosmannia clavigera

AbstractProtocols for the RNA and DNA extractions, cDNA synthesis and qPCR and RT-qPCR for detecting and assessing viability of the Phytophthora ramorum and Grosmannia clavigera tree pathogens., MethodsFifty to 80 mg of wood scrapings were collected at each inoculation point from eight logs of four conifer species and used for the simultaneous extractions of gDNA and mRNA. Using the diluted RNA, cDNA, synthesis was performed using the QuantiTect Reverse Transcription Kit (QIAGEN).