Messenger RNA: What is it? Quality Determination, Intermediary RNA, Classification and Reliable Analysis

It is the ribonucleic acid which transfers the genetic code that comes from the DNA of the cell nucleus to a ribosome in the cytoplasm.

What is RNA?

The RNA research record is quite short. It was not until the first half of the 20th century that RNA was identified as a biological macromolecule independent of DNA by the enterprising chemists of carbohydrates.

Since this distinction was made, multiple classes of RNA have been found, including: messenger RNA (mRNA), ribosomal RNA (rRNA), small RNA, micro RNA and long non-coding RNA.

Similar to DNA, RNA presents a unique set of challenges for researchers, including RNase susceptibility, degradation, and omnipresent shear by pipetting.

The molecular cousin of DNA, RNA plays innumerable important roles in molecular biology, which include: serving as a genome, enzymatic activity and participating in every step of gene expression.

Because of the ubiquity of RNA in biology, scientists have invested time and effort in understanding its chemical and biological properties.

In an effort to understand the biology of RNA, researchers have developed a multitude of relevant molecular biology techniques, ranging from reverse transcription PCR and agarose gel electrophoresis, to micro-chains and next-generation sequencing.

In order to modernize and simplify RNA analysis and quality control workflows, it produces several solutions for RNA workflows.

RNA is important for numerous applications of molecular biology, which include: qPCR, NGS and micro chains. A deep understanding of RNA biology affects a wide range of biological research, from agriculture and medicine to the evolution and editing of the genome.

Ubiquitous in biology, RNA is crucial for diverse and significant processes in molecular biology. As such, it is a biological macromolecule of interest to scientists of diverse origins and interests.

Fundamental to the understanding of everything from the origin of life to the biology of cancer, numerous applications have been developed to better understand the biological functions of RNA.

Sophisticated subsequent applications, especially NGS, require the use of RNA of adequate quality to guarantee reliable results.

However, traditional methods of RNA analysis lack the performance, sensitivity and precision necessary to evaluate with confidence the suitability of RNA for subsequent applications.

For example, RNA agarose gel electrophoresis is a low performance solution that, in addition to lacking adequate resolution and quantification capabilities, is tedious and time consuming.

Total RNA and quality determination

The determination of the total RNA quality is of absolute importance for the analysis of micro chains, NGS and other applications sensitive to RNA degradation and contamination.

Legacy methods of RNA analysis require high input concentrations with limited resolution and quantification capabilities. And they provide a higher quantification, rating and size of the total RNA samples.

To assist in the empirical evaluation of total RNA quality, AATI developed the RNA Quality Number (RQN) for the data analysis software.

Calculated automatically by PROSize for each sample of total RNA, RQN takes into account several factors, including the relationship between ribosomal peaks, the small region of RNA and the resolution between peaks. On a scale of 1 to 10, the highest values ​​indicate a higher total RNA sample.

The DNF-471 standard sensitivity RNA analysis kit and the DNF-472 high sensitivity RNA analysis kit.

The separation resolutions of these kits provide researchers with a clear view of the various separation profiles of total RNA (Download Application Note: “Detection of plant RNA degradation with the fragment analyzer”).

Messenger RNA (intermediary)

The messenger RNA (mRNA), the intermediary between the DNA and the protein, is of interest to the researchers, since it provides information on the activity of the genes.

Large-scale sequencing of an organism’s mRNA, called transcriptomics, allows researchers to determine which genes and genomic regions are active under different conditions.

As with other NGS applications, proper CC analysis of crude nucleic acids ensures that only appropriate samples are sequenced.

Small RNA (classification)

Small RNA is a broad and growing classification, which includes: microRNA, siRNA, piRNA and snRNA. The analysis of small RNAs is often difficult, since many analysis methods lack the minimum resolution and sensitivity required for accurate separation, sizing and quantification.

The DNF-470 small RNA analysis kit for the fragment analyzer and the INFINITY fragment analyzer provides supreme sensitivity, variable performance and remarkable resolution for the analysis of small size RNA.

FFPE RNA and reliable analysis

Reliable CC analysis of FFPE RNA is essential to obtain confidential results of NGS.

FFPE RNA samples exhibit higher levels of degradation and cross-linking, which limits the application of RQN ​​to these samples.

The DV200 quality metric for PROSize was developed by Ilumina scientists to address the unique quality control challenges of FFPE RNA.

The DV200 provides researchers with a powerful tool to reliably analyze FFPE RNA samples suitable for NGS from inadequate samples, saving time and costs.