One step superscript 3 one step rt pcr kit

Viral RNA was extracted from nasal wash samples using the RNeasy mini kit and RNase-free DNase set (Qiagen, Germantown, MD). RNA was reverse transcribed using the one-step SuperScript III one-step RT-PCR kit (Invitrogen, Grand Island, NY) and amplified by segment-specific PCR (primers listed in Table S1) using Phusion high-fidelity DNA polymerase (New England Biolabs, Ipswitch, MA). PCR products were purified from a 1% agarose gel using the QIAquick Gel Extraction Kit (Qiagen, Germantown, MD) and eluted in water. Cleaned PCR product was quantified with the Qubit dsDNA HS Assay Kit (Invitrogen, Grand Island, NY) and diluted in DEPC-treated water to a concentration of 0.2 ng/μl. reverse transcribed, amplified and prepared for sequencing using the Nextera XT Sample Preparation kit. Samples were processed on the Illumina MiSeq with a 500-cycle kit as an 8 pM library with 1% PhiX control.