Near full-length 16S rRNA gene amplicon sequencing of culture KS, originating from a freshwater sediment core
DNA was extracted using the DNEasy UltraClean Microbial Kit (QIAGEN, Hilden, Germany), according to the user's manual. Two rounds of PCR were applied for DNA amplification. The first PCR was performed using the KAPA HiFi ReadyMix PCR Kit (KAPABioSystems, Cape Town, South Africa) and universal primer-tailed 16S primers (27F /5AmMC6/ gcagtcgaacatgtagctgactcaggtcac AGRGTTYGATYMTGGCTCAG, 1492R /5AmMC6/ tggatcacttgtgcaagcatcacatcgtag RGYTACCTTGTTACGACTT) (Biomers.net, Ulm, Germany), followed by purification with the QIAquick PCR purification kit (QIAGEN, Hilden, Germany). The second PCR was performed using the KAPA HiFi ReadyMix PCR Kit and PacBio Barcoded Universal F/R Primers Plate  96 (Pacific Biosciences, California, USA), followed by AMPure PB bead kit (PacBio biosciences, California, USA) purification. The SMRTbell library preparation and further purification were achieved by SMRTbell Template Prep Kit (PacBio biosciences, California, USA).