Virome of fecal samlies from mice in Zhenjiang city, China
In October 2018, in order to study the positive rate of murine norovirus and the type of virus in mouse feces, we collected 39 samples of mouse feces from Jiangsu University to establish a library. All samples were collected by disposable materials. Fecal specimens of mice were grounded using steel balls and re-suspended in ten volumes of phosphate-buffered saline (PBS) and vigorously vortexed for 5 min. The grounded samples were then frozen and thawed three times on dry ice. The supernatants were then collected after centrifugation (10 min, 15,000g). 500 l of each supernatant was filtered through a 0.45-m filter (Millipore) to remove eukaryotic and bacterial cell-sized particles. The filtrates enriched in viral particles were treated with DNase and RNase to digest unprotected nucleic acid at 37C for 60 min. Remaining total nucleic acid was then isolated using QIAamp MinElute Virus Spin Kit (Qiagen) according to manufacturer's protocol. The library was then constructed using Nextera XT DNA Sample Preparation Kit (Illumina) and sequenced using the Illumina Hiseq 2500 platform with 250 bases paired ends with dual barcoding for each pool. For bioinformatics analysis, paired-end reads of 250 bp generated by MiSeq were debarcoded using vendor software from Illumina.
SUB8249863
ratnoro95