Separation of oligo-RNA by reverse-phase HPLC.
AUTOR(ES)
McFarland, G D
RESUMO
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/ammonium acetate pumped at pressures of 1500-300 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 mononucleosides, mononucleotides, and larger oligomers through at least 20-mers. The absolute detection limit is approximately 1 pmole of base though most of the analytical separations described use approximately 1 nmole. In favorable circumstances it is possible to use the analytical colums to purify approximately 1 mg of an oligonucleotide in a single 10-30 min. elution.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=342283Documentos Relacionados
- Two forms of murine epidermal growth factor: rapid separation by using reverse-phase HPLC.
- Separation of bacterial ubiquinones by reverse-phase high-pressure liquid chromatography.
- Efficient computation of optimal oligo–RNA binding
- Reverse-phase HPLC of DNA restriction fragments and ribooligonucleotides on uncoated Kel-F powder.
- Solid-phase extraction of nitro-PAH from aquatic samples and its separation by reverse-phase capillary liquid chromatography