A review on native and denaturing purification methods for non-coding RNA (ncRNA)

J Chromatogr B Analyt Technol Biomed Life Sci. 2019 Jul 1:1120:71-79. doi: 10.1016/j.jchromb.2019.04.034. Epub 2019 Apr 26.

Abstract

Recently, non-coding RNA (ncRNA) became the centerpiece of human genome research. Modern ncRNA-based research has revolutionized disease diagnosis and therapeutics. However, decoding structural/functional information of ncRNA requires large amount of pure RNA, and hence effective RNA preparation and purification protocols. This review focuses on purification schemes of synthetic oligonucleotides, particularly liquid chromatographic (LC) techniques as improved alternatives to urea-polyacrylamide gel electrophoresis (urea-PAGE) purification. Moreover, the review summarizes the shortcomings of urea-PAGE purification method and details the chromatographic purification such as affinity, ion-exchange (IE) or size-exclusion (SE) chromatography. Specifically, we discuss denaturing and native RNA purification schemes with newest developments. In short, the review evaluates nucleic acid purification schemes required for various structural analyses.

Keywords: Affinity purification; FPLC; HPLC; Liquid chromatography; Oligonucleotides purification; Urea-PAGE; ncRNA.

Publication types

  • Review

MeSH terms

  • Chromatography, Liquid / methods*
  • Electrophoresis, Polyacrylamide Gel / methods
  • Oligonucleotides / analysis
  • Oligonucleotides / chemistry
  • Oligonucleotides / isolation & purification
  • RNA, Untranslated* / analysis
  • RNA, Untranslated* / chemistry
  • RNA, Untranslated* / isolation & purification

Substances

  • Oligonucleotides
  • RNA, Untranslated