Faster, safer, and better DNA purification by ultracentrifugation using GelRed stain and development of mismatch oligo DNA for genome walking

Biosci Biotechnol Biochem. 2014;78(11):1902-5. doi: 10.1080/09168451.2014.940831. Epub 2014 Aug 1.

Abstract

Purification of plant DNA involves lengthy ultracentrifugation using ethidium bromide. Here, ultracentrifugation method is improved by staining with GelRed. The resulting method is faster, safer and of higher sensitivity. Purified DNA quality was confirmed by treatment with restriction enzymes and isolation of gene promoters. New type of long adaptor with mismatch sequence was also developed for promoter isolation.

Keywords: DNA extraction; GelRed; ethidium bromide; ultracentrifugation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Protein Complex Subunits / chemistry
  • Base Pair Mismatch*
  • Base Sequence
  • Coloring Agents / chemistry*
  • DNA / isolation & purification*
  • Genomics / methods*
  • Molecular Sequence Data
  • Polymerase Chain Reaction
  • Time Factors
  • Ultracentrifugation

Substances

  • Adaptor Protein Complex Subunits
  • Coloring Agents
  • DNA