GSM7856893: LCA4; Mus musculus; RNA-Seq
Total RNA was extracted from samples using RNA Isolater Total RNA Extraction Reagent (Vazyme, Nanjing, China). The RNA integrity and DNA contamination of the samples were analyzed by agarose gel electrophoresis, and the RNA purity (OD260/280 and OD260/230 ratio) and RNA integrity were determined using a NanoPhotometer spectrophotometer and Agilent 2100 bioanalyzer. The initial RNA for library construction was total RNA with a total amount ≥ 1 μg. NEB common library construction: The first cDNA strand was synthesized in M-MuLV reverse transcriptase system using fragmented mRNA as template and random oligonucleotides as primer, then the RNA strand was degraded by RNaseH, and the second cDNA strand was synthesized using dNTPs as raw material in DNA polymerase I system. The purified double-stranded cDNA was end-repaired, A-tail was added, and sequencing joints were connected. cDNA of about 250-300bp was screened by AMPure XP beads for PCR amplification, and PCR products were purified by AMPure XP beads again, and finally A library was obtained. The NEBNext® Ultra™ RNA Library Prep Kit was used for Illumina®.