16s rRNA profiling: norway rat feces
The preparation of amplicon libraries was carried out according to a two-step protocol, where the first PCR was carried out with degenerate nucleotide primers for the V4 region of the 16s rRNA gene F515 (5`-GTGBCAGCMGCCGCGGTAA-3') and 806R (5'-GGACTACHVGGGTWTCTAAT-3'), supplemented with a "heterogeneous spacer" of 1-5 bp and a linker sequence, and the second PCR introduced Illumina platform-compatible barcodes and adapters to the flow cell [2]. Amplification was carried out with the reaction mixture "qPCRmix-HS SYBR" (Evrogen, Russia) on a thermal cycler "CFX96" (BioRad, CA, USA) in accordance with the works of Toshakov et al.
SUB15589506
17A