High quality DNA from Fungi for long read sequencing e.g. PacBio, Nanopore MinION v4
説明
<p>Extraction of high quality DNA for long read sequencing e.g. PacBio and MinIon Optimized for DNA extraction fromBipolaris sorokiniana. Also tested on Parastaognospora nodorum, Zymoseptoria tritici, wheat stripe rust, barley stripe rust and Pyrenophora tritici-repentis Buffers are best when fresh and not older than 3-6 months. Buffered Phenol:Chloroform:Isoamylalcohol (25:24:1) should not be older than 3 months. Critical steps to obtain high quality DNA: Do NOT heat samples during DNA extractions! Perform all steps at RT or 4oC as indicated. Do NOT incubate samples with KAc for prolonged time periods Perform two steps of buffered Phenol:Chloroform:Isoamylalcohol purification to reduce co-purifying metabolites. DNA fragments were well above the 40kb mark based on Pippin Pulse Gels. The sequencing center performed a second AMPure purification step before library construction. </p>