CD Genomics introduced accurate and affordable RNA-seq and Total RNA sequencing. CD genomics combines proven ribosomal reduction library preparation chemistries and Illumina NGS technology into a single, streamlined protocol, which enable researchers to quickly and easily conduct gene expression studies that provide a complete view of the transcriptome, even from low-quality RNA samples.
“I’ve been working with CD Genomics for many years, every time, the company delivers satisfactory results to me”, said by one of the company’s clients, “Our research runs smoothly when we get a rich source of information on changes in gene expression and regulation associated with cancer and a wide range of other diseases.”
Using RNA-seq or total RNA sequencing?
RNA sequencing (RNA-Seq) is a powerful and comprehensive method for analyzing cell transcriptomes, which enables the qualitative and quantitative detection of multiple RNAs in biological samples at specific points in time. RNA-Seq has a wide range of applications, from basic research on cell structure and function to the detection of various disease states in clinical samples. For example, the difference in gene expression before and after treatment intervention can be compared to determine whether the disease exists. RNA-Seq can also be used to detect alternative splicing patterns, post-transcriptional modifications, and exon-intron boundaries. The obtained data can provide valuable information for basic cellular mechanisms, genomic structure, disease-inducing effects, etc.
Total RNA samples consist of multiple types of transcripts, including coding and non-coding RNA. Messenger RNA (mRNA) is the coding portion of total RNA, accounting for only 1-4% of total RNA, and the rest is usually considered non-coding RNA. Non-coding RNA (ncRNA) refers to RNA that is not translated into protein. These RNAs include ribosomal RNA (rRNA), transport RNA (tRNA), long noncoding RNA (lncRNA), and many smaller ncRNAs such as microRNA (miRNA), Small nuclear RNA (snRNA), small interfering RNA (siRNA), RNA interacting with piwi protein (piRNA), nucleolar small molecule RNA (snoRNA), etc.
When performing RNA sequencing, the choice between total RNA-Seq or mRNA-Seq depends primarily on the purpose of the experiment, and there are some important differences between the two methods. Total RNA-Seq (also known as whole transcriptome sequencing) is the most comprehensive method and usually involves sequencing all RNA molecules, both encoded and non-encoded.
If the research target is eukaryotes and the main focus is on coding regions, mRNA-Seq is a better choice. The mRNA-Seq protocol uses a screening method to enrich poly (A) RNA. mRNA accounts for only a small part of total RNA molecules, so if only sequencing the mRNA can meet the experimental purpose, mRNA-Seq is the most effective and cost-effective method.
Additional consideration should be given to the overall budget before choosing an approach. Total RNA-Seq generates more sequencing data (usually 100-200 million sequencing reads per sample) and costs more than mRNA-Seq. If only mRNA information is needed, mRNA-Seq can provide greater sequencing depth and cost less than total RNA-Seq. This is because sequencing reads (typically 25-50 million sequencing reads per sample) are concentrated on Poly (A)-enriched RNA molecules.
In total RNA-Seq and mRNA-Seq protocols, both ribosome removal and mRNA enrichment methods can improve the quality of sequencing data. In this way, only the target RNA molecules can be sequenced after processing, which minimizes the waste of sequencing reads.
CD Genomics provides both accurate and affordable RNA-seq and Total RNA sequencing, including sample standardization, library construction, deep sequencing, raw data quality control, genome assembly, and customized bioinformatics analysis.