16s-seq of Canis lupus familiaris feces: EcN-U biol repl2
Using 20-50 ng of DNA as a template, a series of PCR primers designed by our company are used to amplify the 16S rDNA of prokaryotes, including the two highly variable regions, V3 and V4. The upstream primer 338F, which contains the sequence "ACTCCTACGGGAGGCAGCAG", and the downstream primer 806R, which contains the sequence "GGACTACHVGGGTWTCTAAT", are used to amplify the V3 and V4 regions (the primers are confidential and not provided in full). Additionally, PCR is used to add adapters with indices to the 16S rDNA PCR product's ends for NGS sequencing. Finally, the PCR product library is analyzed by electrophoresis on a 1.5% agarose gel to verify the target fragment at approximately 600 bp.