A small-scale plasmid preparation yielding DNA suitable for double-stranded sequencing and in vitro transcription

Research output: Contribution to journalArticle

10 Scopus citations

Abstract

A small-scale plasmid preparation is described that is useful for a variety of procedures from double-stranded sequencing to in vitro transcription. No specialized equipment or reagents are required. The preparation of plasmid DNA does not require the use of RNase; instead the larger RNAs are precipitated with 2.5 m ammonium acetate. The resulting plasmid DNA is used routinely for double-stranded sequencing with the Klenow fragment of DNA polymerase and has been used for generating deletions with exonuclease III. In addition, the plasmid DNA has been used to generate transcripts with T7 RNA polymerase that translate well in reticulocyte lysate.

Original languageEnglish (US)
Pages (from-to)170-174
Number of pages5
JournalAnalytical Biochemistry
Volume190
Issue number2
DOIs
StatePublished - Nov 1 1990
Externally publishedYes

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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