DNA extraction was conducted using glass beads (CapitalBio, Beijing, China) and boiling method as earlier reported with a little revision [26 (link)–28 (link)]. Briefly, a bit of sediments of CSF after centrifugation or fresh colonies were suspended in 50 μl of 1× TE buffer (10 mM Tris-HCl, 1 mM EDTA, pH 8.0) within an extraction tube and incubated at 95 °C in a boiling water-bath for 5 min. Then the total DNA was isolated by vortexing at maximum speed in an Extractor™ 36 (CapitalBio) for 5 min. After centrifuged at 10000 g for 5 min, the supernatant of the lysate containing gemonic DNA was separated for follow-up tests. This is “glass beads method” for sample cell lysate.
In order to meet the special needs of Fig.4 , we have further purified the supernatant of the lysate with “TIANamp Yeast DNA Kit” (TIANGEN Biotech, Beijing, China) starting from step 7 of the operating manual. This is “spin columns method” for DNA extraction.
DNA concentrations were accurately measured using the “Qubit™ 3 Fluorometer” (Q33216, Thermo Fisher Scientific, Wilmington, USA). All genomic DNA and sample lysate were kept in − 80 °C to preserve, and avoid repeated freeze-thaw cycles.
In order to meet the special needs of Fig.
DNA concentrations were accurately measured using the “Qubit™ 3 Fluorometer” (Q33216, Thermo Fisher Scientific, Wilmington, USA). All genomic DNA and sample lysate were kept in − 80 °C to preserve, and avoid repeated freeze-thaw cycles.
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