Transcriptome Profiling of Hepatopancreas

Use RNAprep pure Tissue Kit (TianGene, Beijing, China) extraction method to extract RNA from hepatopancreas tissue, and then strictly control the quality of the RNA sample. The quality control method is mainly through the Agilent 2100 bioanalyzer, then the RNA integrity is tested, and the experiment is as follows The NEB common library building method is used to build the library (19 (link)), using fragmented mRNA as a template, random oligonucleotides as primers, and synthesizing the first strand of cDNA in the M-MuLV reverse transcriptase system Second strand cDNA synthesis was subsequently performed using DNA Polymerase I and RNase H. After adenylation of 3′ ends of DNA fragments, NEBNext Adaptor with hairpin loop structure was ligated to prepare for hybridization (20 (link)). To select cDNA fragments of preferentially 250 ~ 300 bp in length, the library fragments were purified with AMPure XP system (Beckman Coulter, Beverly, USA) Then 3 μL USER enzyme (NEB, USA) was used with sizeselected, adaptor-ligated cDNA at 37°C for 15 min followed by 5 min at 95°C before PCR. PCR was performed with Phusion High-Fidelity DNA polymerase, Universal PCR primers, and Index (X) Primer. At last, PCR products were purified (AMPure XP system) and library quality was assessed on the Agilent Bioanalyzer 2100 system (8 (link), 21 (link)).