High-speed preparative-scale separation and purification of ribosomal 5S and 5.8S RNA'S via sephacryl S-300 gel filtration chromatography

Kai Mon Lee, Alan G. Marshall

Research output: Contribution to journalArticlepeer-review

5 Scopus citations

Abstract

In this work we present a rapid and economical alternative to Sephadex and high performance size exclusion chromatography (HPSEC) for preparative-scale separation and purification of low molecular weight RNA's: 5.8S RNA, 5S RNA, and tRNA's. These three RNA species can be well resolved from each other and from higher molecular weight RNA species via Sephacryl S-300 gel filtration chromatography under mild eluting conditions: 10 mM Tris-HCl buffer, pH 7.5, containing 1.0 M NaCl. For a sample load of about 250 mg, the resolving power of a Sephacryl S-300 column (78 × 3.2 cm) is comparable to that of a 4.5 times larger Sephadex G-75 column (144 × 5 cm). Moreover, the total separation period is 2.5 times shorter for the Sephacryl method. Up to 500 mg or more of crude ribosomal RNA mixtures could be separated via two Sephacryl S-300 columns operated in tandem.

Original languageEnglish (US)
Pages (from-to)247-258
Number of pages12
JournalPreparative Biochemistry
Volume16
Issue number3
DOIs
StatePublished - Sep 1 1986
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Genetics

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