Agilent 2100 biological

The RNeasy mini kits were used to isolate the RNA, and strand-specific libraries were prepared using the VAHTS Total RNA-seq (H/M/R) Library Prep Kit (Vazyme, China). In Brief, RNA was purified after removal of rRNA by magnetic beads. The cleaved RNA fragments, which were segmented with divalent cations(94°C, 8 min), were used to synthesize the first strand cDNA; then, the reverse transcriptase and random primers were used to generate the second strand cDNA. After PCR purification and enrichment, the final cDNA library was generated. Then, Qubit® 2.0 Fluorometer (Life Technologies, USA) was used for quantification, and Agilent 2100 biological (Agilent Technologies, USA) was used for verification, so as to determine the size of the insert and calculate the molar concentration. cBot was used for cluster analysis, and Illumina NovaSeq 6000 (Illumina, USA) was sequenced. Shanghai Sinomics Corporation constructed and sequenced the library.

Total RNA from SL0 and L3 hepatic tissues was extracted using a Trizol kit (Invitrogen, Carlsbad, CA, USA). The total RNA quality and integrity were examined using an Agilent 2100 biological analyzer (Agilent Technologies, Palo Alto, CA, USA) and RNase-free agarose gel electrophoresis, respectively. The library construction and high-throughput sequencing were completed by Genedenovo Biotechnology Co., Ltd. (Guangzhou, China), and high-throughput sequencing was performed using an Illumina NovaSeq 6000.

Total RNA was extracted using the Qiagen RNeasy kit (QIAGEN), and during the purification process of the sample, DNA enzyme (QIAGEN) was used for on-column digestion of the DNA. RNA quality and concentration were measured using an Agilent 2100 biological analyzer.