In situ labeling of DNA breaks and apoptosis by T7 DNA polymerase

Methods Mol Biol. 2011;682:37-48. doi: 10.1007/978-1-60327-409-8_4.

Abstract

The native T7 DNA polymerase is a fast and highly processive enzyme that can be used for in situ detection of apoptosis and various types of DNA breaks. The technique is quick and simple, and was shown to label earlier stages of apoptosis compared to the terminal transferase technique. The in situ labeling applications of T7 DNA polymerase are presented and summarized from the DNA damage detection standpoint. The detailed protocols are provided together with the discussion of their advantages and limitations.

MeSH terms

  • Animals
  • Apoptosis*
  • DNA Breaks*
  • DNA-Directed DNA Polymerase / metabolism*
  • Formaldehyde
  • Frozen Sections
  • In Situ Nick-End Labeling / methods*
  • Paraffin Embedding
  • Tissue Fixation

Substances

  • Formaldehyde
  • bacteriophage T7 induced DNA polymerase
  • DNA-Directed DNA Polymerase