title: Illumina NovaSeq 6000 paired end sequencing; Characterization of the transcriptome profile of bursa of Fabricius infected by Muscovy duck reovirus in muscovy ducklings
design: Characterization of the transcriptome profile of bursa of Fabricius infected by Muscovy duck reovirus in muscovy ducklings
library_name: HN10_1_lncRNA_p
library_construction_protocol: Twenty seven 1-day-old healthy Muscovy ducklings were collected and randomly separated into three groups. Nine ducklings were administrated with 500 μL JDm10 strain at a titer of 108.2 median tissue culture infective dose (TCID50) per mL via intramuscular injection; nine ducklings were administrated with 500 μL HN10 strain at a titer of 106.4 median TCID50 per mL; the rest of ducklings were treated with sterile DMEM as a blank control. The bursa of Fabricius specimens were harvested at 72 hours post infection (hpi) and frozen in liquid nitrogen for the consequent experiments. Total RNA of bursa of Fabricius tissues was extracted using RNAiso Plus (Takara Bio Inc., Dalian, China). The RNA amount and purity of each sample was quantified using NanoDrop ND-1000 (NanoDrop, Wilmington, DE, USA). The RNA integrity was assessed by Bioanalyzer 2100 (Agilent, CA, USA) with RIN number >7.0, and confirmed by electrophoresis with denaturing agarose gel. RNA was divided into two parts for at least 2 μg in each. Epicentre Ribo-Zero Gold Kit (Illumina, San Diego, USA) was used to deplete rRNA, and NEB Next Ultra Directional RNA Library Prep Kit for Illumina (New England Biolabs, Ipswich, USA) was used to establish the library of one part for long chain RNA sequencing. TruSeq Small RNA Sample Prep Kit (Illumina) was used to establish the library of the other part for small RNA sequencing.