In vitro transcription from plasmid or PCR-amplified DNA

Julie L. Brunelle, Rachel Green

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

This protocol describes the synthesis and purification of RNAs using plasmid DNA or PCR-amplified DNA as a template. This procedure should give NTP-free, full-length RNA for all sizes of RNA. This protocol is derived from Milligan and Uhlenbeck, the classic paper on T7 transcription reactions, with modifications.

Original languageEnglish (US)
Title of host publicationLaboratory Methods in Enzymology
Subtitle of host publicationRNA
PublisherAcademic Press Inc.
Pages101-114
Number of pages14
ISBN (Print)9780124200371
DOIs
StatePublished - 2013

Publication series

NameMethods in Enzymology
Volume530
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988

Keywords

  • Column purification
  • DNA
  • DNA Phenol extract
  • In vitro transcription reaction
  • PCR-amplified
  • RNA polymerase
  • Template

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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