Mag bind stool dna kit

In total, 270 male yellow broilers were raised at the breeding farm of Jiangsu Xingmu Agricultural Science and Technology Co. (Beijing, China). Each chicken was raised in an independent cage with a food conditioner in the same environment from hatching to 63 days. The feeding experiment was divided into three stages: from day 1 to 20, from day 21 to 40, and from day 41 to 63. The feed formula (Supplementary Table 1) was the same as that described by Huang et al. (2021) (link) and met the nutritional requirements set forth in the Nutritional Requirements of Chickens (1994). The feed intake and BW per chicken were measured every 5 days. The FCR was calculated as the ratio of FE to BWG during days 5 to 63 of feeding. According to the FCR ranking, 22 chickens with high FCR were assigned to the high FE (HFE) group and 22 chickens with low FCR were assigned to the low FE (LFE) group.
On day 64, chickens from each group were euthanized, and samples of the duodenum and ileum were aseptically collected. The samples were stored at −80°C. Microbial genomic DNA was extracted and purified using a Mag-Bind Stool DNA Kit (Omega Bio-Tek, Norcross, GA, United States) according to the manufacturer’s instructions. The quality and quantity of the purified DNA were verified using a NanoDrop spectrometer (Thermo Fisher Scientific, Waltham, MA, United States).

Genomic DNA from rumen and fecal samples was extracted using the Mag-Bind Stool DNA Kit (Omega Bio-Tek, Norcross, GA, U.S.), according to the manufacturer’s protocol with a minor modification. For the lysing step, the protocol was adapted to use a mixer mill (Retsch MM 200) set up for 10 min at 25 Hz. The quality of the resulting DNA was checked using gel electrophoresis (1% agarose gel) and the concentration was measured with a NanoDrop One spectrophotometer (Thermo Fisher Scientific, Wilmington, DE, USA). Bacterial amplicon libraries of the V4 region from the 16S rRNA gene were prepared as described by Paz et al. [20 (link)]. The resulting amplicons were normalized to a concentration of 1–2 ng/μl using the SequalPrep^TM normalization plate. Pooled libraries were sequenced using the Illumina MiSeq platform (Illumina, San Diego, CA, USA). Raw sequences are available at the NCBI Sequence Read Archive (SRA) under the accession no. SRP271418.