There are several challenges in RNA extraction, from RNA degradation and low yield to DNA contamination. Different sample types may require different treatments. While tough cell walls are refractory to chemical and enzymatic lysis, plant samples may contain inhibitors that can co-precipitate with RNA.
Read on for some of the best RNA isolation tips from experts to recover high-quality, DNA-free RNA.
Stabilizing RNA after collection
RNA is highly vulnerable to degradation, especially in samples containing high levels of RNases. This is why it is important to stabilize samples at the time of collection. Common sample stabilization methods include–
- snap freezing with liquid nitrogen
- dry-ice ethanol baths
- storage in a -80°C freezer
But it is important to remember that these approaches carry risks, such as freeze-thaw damage of nucleic acids.
A safer method of RNA Stabilization is immediate solubilization in a lysis buffer to inactivate RNases. Once that is done, the samples can be processed or stored frozen.
The other option is submersion in a stabilization reagent to inactivate nucleases and protect nucleic acids at ambient temperature for longer periods of time. This is best for researchers, who are collecting samples in the field.
Ensure complete sample lysis
You can maximize RNA yield and quality with complete sample lysis. However, not all samples can undergo the same lysis regimen. For example, adding a detergent-based lysis buffer may not always be sufficient for blood cells and microbial cells. In such cases, it is better to pair the lysis buffer with mechanical lysis or add an enzymatic lysis step upstream.
Incomplete lysis can cause buffer carryover, column clogging, DNA contamination, and incomplete elution. Complete sample lysis also aids the extraction procedure too. Considering these factors, Magbio Genomics has developed the High Prep® Viral RNA kit that contains the Viral Lysis Buffer, RDW Buffer, Nuclease-Free Water, Pro K Solution, NBE, LES and MAG-S1 Particles for rapid and reliable isolation of viral nucleic acids from serum, plasma, saliva and other body fluids as well as nasopharyngeal swabs
Eliminate DNA contamination
DNA contamination can skew-based quantification methods, inflating the RNA quantification above actual levels. It can also give false readings in sensitive downstream applications, like RNA sequencing. Ensure accurate quantification measurements and avoid discrepancies in downstream analysis by eliminating DNA carryover. This can be achieved with DNA removal columns and DNase treatment.
The fastest method to detect DNA is to visualize the RNA samples and look for any high molecular weight fragments. qPCR or Qubit can also be used in downstream applications with sensitivity to DNA contamination.
Select a High-Quality RNA extraction kit
High-quality RNA extraction is possible from a variety of samples by ensuring –
- sample stabilization post collection
- complete lysis of the sample
- eliminating DNA contamination
Taking these three steps can make it easier to recover RNA for various downstream applications and accurate results. All-inclusive products, like the High Prep® Viral RNA kit can simplify the extraction process to a great extent.
Magbio Genomics has developed the High Prep® RNA extraction kits. Its LES or Lysis Enhancing Solution ensures optimal lysis of samples, guaranteeing high yields and consistent Ct values. It is highly suitable for the following facilities –
- SAR-CoV-2 Testing & Research Labs
- Diagnostic Labs
- Sequencing Cores working to sequence viral genomes
- Genotyping facilities
- Medical / Clinical Laboratories
- Academic Genomic Cores doing viral research, detection, or testing
- Biotech / Pharma companies developing viral detection panels/assays
- Hospitals monitoring patient viral load
If you are looking for viral DNA/RNA extraction kit and RNA extraction kit manufacturers in the USA, call MAGBIO Genomics at 301-302-0144. MAGBIO Genomics is the industry leader in magnetic bead-based products for nucleic acid isolation and DNA, cfDNA, and RNA extraction.