RNA-Seq of Neural tube organoid
Total RNA was extracted using the TRIzol reagent (Invitrogen, CA, USA). VAHTS Universal V6 RNA-seq Library Prep Kit was used for preparation of the RNA-seq library. RNA-seq was performed using the llumina Novaseq 6000 platform and 150 bp paired-end reads were generated. Raw reads of fastq format were firstly processed with fastp1 for Illumina adapter trimming and removal of low-quality reads. For gene expression analysis, sequence FASTQ files were mapped to the reference human genome using HISAT2. FPKM (fragments per kilobase of the exon model per million mapped fragments) of each gene was calculated and the read counts of each gene were obtained by HTSeq-count. The transcriptome sequencing and analysis were conducted by OE Biotech Co., Ltd. (Shanghai, China).