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New App Note: RP analysis of RNA markers using bioinert YMC-Accura Triart Bio C4
Today, oligonucleotides have become more and more important in genetic testing, research, and forensics.
To date a variety of oligonucleotide-based approaches have been developed for different pharmaceutical applications. Therefore, clean and reliable oligonucleotide standards are necessary for various analytical methods. One of these standards are RNA markers which are RNA fragments of a certain size which are used in gel electrophoresis to estimate the size of other RNA fragments.
The electron-rich backbone of oligonucleotides can cause irreversible adsorption on metal parts. This is a major issue for reversed phase liquid chromatography analyses as standard equipment and hardware such as tubing, frits or the standard column-body is made from stainless steel. The oligonucleotides bind through ionic interactions with the positively charged metal oxide layer of the metallic surfaces. This effect is enhanced when working at low to neutral pH as metals become more electropositive. To overcome these challenges bioinert systems and columns such as the recently introduced YMC-Accura Triart columns are beneficial. YMC-Accura Triart columns have a bioinert coating on all surfaces, including the frits, to prevent any unwanted ionic interactions.
This new application note describes the analysis of 7 RNA markers (100 – 1000 bases) using the recently introduced bioinert YMC-Accura Triart Bio C4 column at pH 7 and 80 °C. While conventional column hardware leads to peak tailing and loss of recovery when analysing oligonucleotides, the bioinert coated YMC-Accura Triart Bio C4 column shows excellent recoveries and peak shapes for all 7 well resolved markers.