Separation of oligo-RNA by reverse-phase HPLC

Gloria Mcfarland, Philip N. Borer

Research output: Contribution to journalArticle

60 Citations (Scopus)

Abstract

A rapid and highly reproducible chromatographic technique has been developed for analysis and purification of complex mixtures of oligoribonucleotides. The method utilizes a column of microparticulate porous silica beads fully derivatized with octadecylsilyl groups. The column is eluted with gradients in acetonitrile/water/anmonium acetate pumped at pressures of 1500-3000 psi. Most separations are completed in 5-15 min. with usually better than 1% reproducibility of absolute retention times and about 0.1% reproducibility of relative retention times. A single column accomplishes separations of menonucleosides, mononucleotides, and larger oligomers through at least 20-mers. The absolute detection limit is ∼1 pmole of base though most of the analytical separations described use ∼1 nmole. In favorable circumstances it is possible to use the analytical columns to purify ∼1 mg of an oligonucleotide in a single 10-30 min. elution.

Original languageEnglish (US)
Pages (from-to)1067-1080
Number of pages14
JournalNucleic Acids Research
Volume7
Issue number4
DOIs
StatePublished - Oct 25 1979

Fingerprint

High-performance Liquid Chromatography
RNA
Reverse
High Pressure Liquid Chromatography
Oligoribonucleotides
Reproducibility
Complex Mixtures
Oligonucleotides
Silicon Dioxide
Limit of Detection
Acetates
Acetonitrile
Oligomers
Pressure
Detection Limit
Purification
Water
Silica
Gradient
High-performance liquid chromatography

ASJC Scopus subject areas

  • Genetics
  • Statistics, Probability and Uncertainty
  • Health, Toxicology and Mutagenesis
  • Applied Mathematics
  • Genetics(clinical)
  • Toxicology

Cite this

Separation of oligo-RNA by reverse-phase HPLC. / Mcfarland, Gloria; Borer, Philip N.

In: Nucleic Acids Research, Vol. 7, No. 4, 25.10.1979, p. 1067-1080.

Research output: Contribution to journalArticle

Mcfarland, Gloria ; Borer, Philip N. / Separation of oligo-RNA by reverse-phase HPLC. In: Nucleic Acids Research. 1979 ; Vol. 7, No. 4. pp. 1067-1080.
@article{92a9aa05ca03483a8b01ab8e298528a3,
title = "Separation of oligo-RNA by reverse-phase HPLC",
abstract = "A rapid and highly reproducible chromatographic technique has been developed for analysis and purification of complex mixtures of oligoribonucleotides. The method utilizes a column of microparticulate porous silica beads fully derivatized with octadecylsilyl groups. The column is eluted with gradients in acetonitrile/water/anmonium acetate pumped at pressures of 1500-3000 psi. Most separations are completed in 5-15 min. with usually better than 1{\%} reproducibility of absolute retention times and about 0.1{\%} reproducibility of relative retention times. A single column accomplishes separations of menonucleosides, mononucleotides, and larger oligomers through at least 20-mers. The absolute detection limit is ∼1 pmole of base though most of the analytical separations described use ∼1 nmole. In favorable circumstances it is possible to use the analytical columns to purify ∼1 mg of an oligonucleotide in a single 10-30 min. elution.",
author = "Gloria Mcfarland and Borer, {Philip N.}",
year = "1979",
month = "10",
day = "25",
doi = "10.1093/nar/7.4.1067",
language = "English (US)",
volume = "7",
pages = "1067--1080",
journal = "Nucleic Acids Research",
issn = "0305-1048",
publisher = "Oxford University Press",
number = "4",

}

TY - JOUR

T1 - Separation of oligo-RNA by reverse-phase HPLC

AU - Mcfarland, Gloria

AU - Borer, Philip N.

PY - 1979/10/25

Y1 - 1979/10/25

N2 - A rapid and highly reproducible chromatographic technique has been developed for analysis and purification of complex mixtures of oligoribonucleotides. The method utilizes a column of microparticulate porous silica beads fully derivatized with octadecylsilyl groups. The column is eluted with gradients in acetonitrile/water/anmonium acetate pumped at pressures of 1500-3000 psi. Most separations are completed in 5-15 min. with usually better than 1% reproducibility of absolute retention times and about 0.1% reproducibility of relative retention times. A single column accomplishes separations of menonucleosides, mononucleotides, and larger oligomers through at least 20-mers. The absolute detection limit is ∼1 pmole of base though most of the analytical separations described use ∼1 nmole. In favorable circumstances it is possible to use the analytical columns to purify ∼1 mg of an oligonucleotide in a single 10-30 min. elution.

AB - A rapid and highly reproducible chromatographic technique has been developed for analysis and purification of complex mixtures of oligoribonucleotides. The method utilizes a column of microparticulate porous silica beads fully derivatized with octadecylsilyl groups. The column is eluted with gradients in acetonitrile/water/anmonium acetate pumped at pressures of 1500-3000 psi. Most separations are completed in 5-15 min. with usually better than 1% reproducibility of absolute retention times and about 0.1% reproducibility of relative retention times. A single column accomplishes separations of menonucleosides, mononucleotides, and larger oligomers through at least 20-mers. The absolute detection limit is ∼1 pmole of base though most of the analytical separations described use ∼1 nmole. In favorable circumstances it is possible to use the analytical columns to purify ∼1 mg of an oligonucleotide in a single 10-30 min. elution.

UR - http://www.scopus.com/inward/record.url?scp=0018801456&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0018801456&partnerID=8YFLogxK

U2 - 10.1093/nar/7.4.1067

DO - 10.1093/nar/7.4.1067

M3 - Article

C2 - 503849

AN - SCOPUS:0018801456

VL - 7

SP - 1067

EP - 1080

JO - Nucleic Acids Research

JF - Nucleic Acids Research

SN - 0305-1048

IS - 4

ER -