Snapshot blotting: Transfer of nucleic acids and nucleoprotein complexes from electrophoresis gels to grids for electron microscopy

S. D. Jett, D. G. Bear

Research output: Contribution to journalArticlepeer-review

5 Scopus citations

Abstract

We present a technique, 'snapshot blotting,' for the electrophoretic transfer of nucleic acids and nucleoprotein complexes in gel electrophoresis bands onto highly stable carbon film-coated grids for imaging by electron microscopy. The method permits structural analysis of macromolecular species that have been resolved by a gel mobility-shift assay. To demonstrate the efficiency and integrity of the transfer process for a multiprotein-DNA assembly, we have imaged various species of a prokaryotic transcription complex, using the cleavage-defective EcoRI(Q111) protein as an orientation marker and as a blockade of transcription elongation. Snapshot blotting should be of great utility in the structural characterization of nucleic acids and protein-nucleic acid interactions.

Original languageEnglish (US)
Pages (from-to)6870-6874
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume91
Issue number15
DOIs
StatePublished - Jul 19 1994
Externally publishedYes

Keywords

  • gel mobility-shift assay
  • protein-nucleic acid interactions
  • transcription

ASJC Scopus subject areas

  • General

Fingerprint

Dive into the research topics of 'Snapshot blotting: Transfer of nucleic acids and nucleoprotein complexes from electrophoresis gels to grids for electron microscopy'. Together they form a unique fingerprint.

Cite this