Nebnext small rna library prep set for multiplex compatible kit

Trizol reagent (Tiangen, Beijing, China) was used to extract total RNA from undifferentiated and differentiated chicken preadipocytes (Add 1 mL of Trizol per 2.5 × 10⁶ cells). The NEBNext Poly(A) mRNA Magnetic Isolation Module kit (New England Biolabs, Ipswich, MA, USA) and NEBNext Small RNA Library Prep Set for Illumina (Multiplex Compatible) kit (New England Biolabs, Ipswich, MA, USA) were used to construct the lncRNA libraries and miRNA libraries following the manufacturer’s protocol, respectively. Six lncRNA libraries (three for each group) and six miRNA libraries (three for each group) were constructed. The 12 constructed libraries were sequenced using the Illumina HiSeqTM 4000 sequencing platform (Illumina Inc., San Diego, CA, USA).

In total, 16 libraries were constructed individually from zebrafish gonads. Library preparation was performed by NEBNext® Small RNA Library Prep Set for Illumina® (Multiplex Compatible) kit following manufacturer’s instructions. Sequencing (1 × 50, v4, HiSeq) was performed at single-end mode with a read length of 50 bp at the Genomics Unit of the Centre for Genomic Regulation (CRG) in Barcelona.