title: The microbiota of farm house chicken’ feces is more diverse than that in poultry-farm and the aerobic cultivatable tetracycline resistant bacteria per contra
design: To ensure that there was enough 16S rDNA V3 hypervariable region in PCR products to harvest, two-step PCR strategy was used. The PCR enrichment of the 16S rDNA V3 hypervariable region was performed with forward primer 5’-XXXXXXXX-TACGGGAGGCAGCAG-3’ and reverse primer 5’-XXXXXXXX-ATTACCGCGGCTGCTGG-3’. The 5’ terminal of every primer contained an 8-base-oligonucleotide tag (before the hyphen), while the sequence after the hyphen was able to pare the sequences of the V3 end region. The detail refer to (54). After the PCR reaction, the electrophoresis was immediately performed to isolate the enriched V3 region DNA fragments from the reaction mixture with loading buffer and SybGreen. All of the products were approximately 200 bp in length and were harvested using a gel extraction kit (Gel Extraction Kit, OMEGA Bio-Tek, USA) following the manufacture’s instructions. All of the enriched DNA was mixed to establish the library (50 ng/sample).
library_name: microbiota