Maxwell rsc cell dna purification kit

Genomic DNA was extracted from the bronchial fluid of NSCLC patients, which was preserved in PreservCyt solution using the Maxwell RSC Cell DNA purification kit according to the manufacturer’s instruction (Promega, Madison, WI, USA). The quantity and quality of purified DNA were evaluated using Nanodrop and Qubit and stored at 4 °C until tested by Cancer hotspot NGS [22 (link),23 (link)]. Direct smears were prepared from residual bronchial fluid, which were either diff-Quick or Papanicolaou (Pap)-stained. Selected diff-Quick and Pap-stained specimens were used for RNA extraction [24 (link)]. Total RNA was extracted using the Maxwell RSC RNA FFPE kit (Promega, Madison, WI, USA) from the smears to use in the RNA-based assay. The quantity and quality of total nucleic acid were evaluated using Nanodrop and Qubit and stored at −70 °C until tested.

For the Cascade/Cas3 activator templates, DNA fragments of hEMX1, mTyr and IAV variants (which include a target site) were designed and purchased from IDT (Table S2). Total mouse genomic DNA from a C57BL/6 strain was used after purification (Maxwell RSC Cell DNA Purification Kit; Promega, Madison, Wisconsin). LAMP SARS-CoV-2 primers were designed against regions of the N gene using PrimerExplorer v.5 (Eiken Chemical Co.; https://primerexplorer.jp/). The primers used for isothermal PCR are listed in Table S3.
Viral RNAs from SARS-CoV-2 were prepared according to the established protocol from the National Institute of Infectious Diseases in Japan (Matsuyama et al., 2020 (link)). Viral RNAs were purified from an infected TMPRSS2-expressing VeroE6 cell line using the QIAamp Viral RNA Mini Kit (QIAGEN) according to the manufacturer’s protocol.