title: 16S rRNA Amplicon-Seq of Kodagu Apis cerana
design: Metagenome sequencing of honey bees (Apis cerana) from two southern regions, Karnataka, India - Shivamoga and Kodagu. In detail, the universal 16S rRNA full-length primers (16F:5-TTTCTGTTGGTGCTGATATTGC-3 and 16R:5-ACTTGCCTGTCGCTCTCTCTTC-3) and Ex-Taq DNA polymerase (Takara) were used forthe amplification of the V1-V9 region using the nucleic acid extracted from gut homogenate of honey bees. The resulting ~1.5kb amplicons were purified using 1.6XAmpure XP beads (Beckmann Coulter, USA) and the same was quantified using a Qubit dsDNAHS assay kit (Cat no. Q32854).The amplicon ends were polished and A-tailed (NEBnext ultra II end repair kit, New EnglandBiolabs, MA, USA). The Blunt TA ligase master mix (M0367L) barcodedthe prepared samples. The library was prepared by ligating sequencing adapters (SQK-NBD114)onto double-stranded DNA fragments using NEB Quick T4 DNA Ligase (New England Biolabs,MA, USA). After purification with AMPure XP beads,the library was checked with Qubit forquantification and was pooled and sequenced on Nanopore MinION (flow cell FLO-FLG111).
identifiers.namespace: SUB14795034
identifiers.id: S4_R2