Nanodrop one uv vis

At day 38, 72 birds (6 birds per replicate) were selected at random from each house and samples were collected from caecum, ileum, and ileum mucosa. The DNA was extracted using a DNA mini spin column (Enzymax LLC, CAT#EZC101, Lexington, KY, USA). The concentration and quality of extracted DNA were measured using a NanoDrop One UV-Vis spectrophotometer (ThermoFisher Scientific, Waltham, MA, USA). The following primers specific to the V3-V4 region of the 16S rRNA gene were used with spacers, barcodes, and Illumina sequencing linkers [34 (link)]. Pro341F (5′-CCTACGGGNBGCASCAG-3′) was the forward primer, and 805R (5′-GACTACNVGGGTATCTAATCC-3′) was the reverse. The 16S amplicon library was then purified using AMPure XP kits (Beckman Coulter, Brea, CA, USA) and sequenced using Illumina MiSeq platform 2 × 250 bp paired-ended configuration.

Samples were collected, and high molecular weight genomic DNA was prepared by the CTAB method followed by purification with the QIAGEN® Genomic kit (Cat#13343, QIAGEN) for regular sequencing, according to the standard operating procedure provided by the manufacturer. DNA degradation and contamination of the extracted DNA was monitored on 1% agarose gels. DNA purity was then assessed using a NanoDrop One UV–VIS spectrophotometer (Thermo Fisher Scientific, USA), of which OD 260/280 ranged from 1.8 to 2.0 and OD 260/230 from 2.0 to 2.2. DNA concentration was subsequently measured using a Qubit 4.0 fluorometer (Invitrogen, USA).