Index
The ribonucleic acid transfers the genetic code from the cell nucleus’s DNA to a ribosome in the cytoplasm.
What is RNA?
The RNA research record is relatively 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.
Like DNA, RNA presents unique challenges for researchers, including RNase susceptibility, degradation, and omnipresent shear by pipetting.
The molecular cousin of DNA, RNA, plays numerous essential roles in molecular biology, including 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.
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 essential for numerous molecular biology applications, including 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. It is a biological macromolecule of interest to scientists of diverse origins and interests.
Fundamental to understanding everything from the origin of life to the biology of cancer, numerous applications have been developed to understand the biological functions of RNA better.
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
Determining the total RNA quality is extremely important for analyzing 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. Moreover, they provide a higher quantification, rating, and size of the total RNA samples.
I developed the RNA Quality Number (QR) for the data analysis software to assist in empirically evaluating total RNA quality.
Calculated automatically by PROSize for each sample of total RNA, RQN considers 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 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 tricky since many 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 provide maximum sensitivity, variable performance, and excellent 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.
Illumina scientists developed the DV200 quality metric for PROSize to address the unique quality control challenges of FFPE RNA.
