One-step enzymatic purification of PCR products for direct sequencing

Curr Protoc Hum Genet. 2001 Nov:Chapter 11:Unit 11.6. doi: 10.1002/0471142905.hg1106s30.

Abstract

The polymerase chain reaction (PCR) has become a powerful and widespread method for analyzing DNA. This protocol offers convenient enzymatic purification of PCR products for direct sequencing without the need for further subcloning, precipitation, or column purification. All reactions may be performed in 96-well PCR plates and are compatible with large-scale sequencing. This method is designed to be convenient and rapid. It is suitable for a variety of PCR templates, including plasmid, lambda, and genomic DNA.

MeSH terms

  • Alkaline Phosphatase
  • DNA / genetics
  • DNA / isolation & purification
  • Exodeoxyribonucleases
  • Genetics, Medical
  • Genetics, Microbial / methods
  • Humans
  • Polymerase Chain Reaction / methods*
  • Sequence Analysis, DNA / methods*

Substances

  • DNA
  • Exodeoxyribonucleases
  • exodeoxyribonuclease I
  • Alkaline Phosphatase