Direct sequencing of long polymerase chain reaction fragments

Mol Biotechnol. 1998 Oct;10(2):183-5. doi: 10.1007/BF02760864.

Abstract

Direct sequencing of polymerase chain reaction (PCR)-generated templates is a commonly used technique in molecular biology laboratories. We describe an improved method for direct sequencing of PCR fragments longer than 20 kb obtained with a commercial mixture of Taq and Pwo DNA polymerases. The sequencing protocol was optimized for an automated infrared DNA sequencer, consistently yielding long reads (500-600 bases).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA / chemistry*
  • DNA / genetics
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / genetics
  • Escherichia coli / chemistry
  • Escherichia coli / genetics
  • Polymerase Chain Reaction
  • Reproducibility of Results
  • Sequence Analysis, DNA / methods*
  • Streptococcus pneumoniae / chemistry
  • Streptococcus pneumoniae / genetics

Substances

  • DNA, Bacterial
  • DNA