mRNA sequencing analyzes the transcriptome, revealing gene expression patterns and novel transcripts in cells or tissues. The process involves RNA isolation, cDNA synthesis via reverse transcription, library preparation with added adapters, high-throughput sequencing, and bioinformatic analysis mapping reads to a reference genome or transcriptome to discern gene expression levels and novel transcripts.
mRNA sequencing from standard RNA input is performed with Illumina Stranded mRNA Prep protocol (Illumina) that ensures precise strand orientation measurement, uniform coverage, and high-confidence detection of novel features like isoforms and gene fusions.
mRNA sequencing from RNA low input is performed with the SMART-Seq v4 PLUS Kit (Takara). SMART technology ensures full-length transcript information, enabling analysis of isoforms, gene fusions, and mutations, with improved gene detection via locked nucleic acid (LNA) technology. High reproducibility and accurate coverage of GC-rich transcripts are ensured.