Initial clinical diagnosis was GO
1.RNA extraction: Isolation of RNA from the biological sample of interest using appropriate methods and reagents.2.RNA fragmentation: The RNA is fragmented into smaller pieces, typically by using divalent cations under elevated temperature.3.cDNA synthesis: Reverse transcription of the fragmented RNA into cDNA using reverse transcriptase and random primers.4.End repair and A-tailing: The cDNA ends are repaired and adenylated to prepare them for adapter ligation.5.Adapter ligation: Unique adapters are ligated to the cDNA fragments to enable sequencing on the BGISEQ500 platform.6.PCR amplification: Amplification of the cDNA fragments with ligated adapters to enrich the library for sequencing.7.Quality control: Assessment of the library quality through methods such as qPCR, bioanalyzer, or TapeStation to ensure it meets the requirements for sequencing.