One-step RNA extraction for RT-qPCR detection of 2019-nCoV

Monica Sentmanat; Evguenia Kouranova; Xiaoxia Cui

We sought to compare yields from direct input of QE buffer-lysed sample and column purified RNA. An OP specimen was placed in 200 ul of QE buffer and the collection swab removed. We removed 20 ul for our experimental input before adding 100 copies/ul of positive control DNA template to the remaining buffer. We took 120 ul for column purification using the Qiagen RNeasy Mini Kit with a final elution volume of 30 ul. Although this is not one of the CDC recommended RNA extraction kits, it has been previously shown to perform as well as the recommended kits for positive control samples 7 . The remaining 60 ul of QE sample positive control counterpart and 20 ul of experimental QE sample were heat extracted. Each RT-qPCR reaction had 2 ul of input assayed. We found QE processed and column purified material had comparable Ct values despite 4-fold more material processed for column purification ( Figure 2 ).