Total RNA sequencing comprehensively analyzes the transcriptome, revealing mRNA and non-coding RNA profiles. This technique aids in understanding gene expression, identifying novel transcripts, and deciphering regulatory mechanisms in various biological processes. The process involves RNA extraction, ribosomal RNA depletion, cDNA library generation via reverse transcription, fragmentation, adaptor ligation, PCR amplification, high-throughput sequencing, and bioinformatic analysis for gene expression quantification and transcript discovery.
Libraries are prepared using the Illumina Total RNA Prep protocol (Illumina) with Ribo-Zero Plus that supports diverse RNA inputs, including FFPE and low-quality samples, and incorporates Ribo-Zero Plus or Ribo-Zero Plus Microbiome for efficient removal of abundant RNA from various species, including human, mouse, rat, bacteria, and complex microbial samples like stool, ideal for meta-transcriptomic studies.