Immunoprecipitation of DNA:RNA Hybrids Using the S9.6 Antibody

Methods Mol Biol. 2020:2161:195-207. doi: 10.1007/978-1-0716-0680-3_14.

Abstract

Formation of DNA:RNA hybrids or R-loops contributes to numerous biologic processes. The development of the S9.6 antibody makes the analysis of R-Loops (DNA:RNA hybrids) possible through immunoprecipitation. Here, we describe the isolation of DNA:RNA hybrid structures using the S9.6 antibody. Using this protocol, both the DNA and RNA binding partners of the R-loop can be analyzed via qPCR, whole genome sequencing, or other methods.

Keywords: ChIP; DNA; DRIP; Immunoprecipitation; R-loop; RNA; S9.6.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Antibodies / immunology*
  • DNA / chemistry
  • HEK293 Cells
  • Humans
  • Immunoprecipitation / methods*
  • R-Loop Structures / immunology*
  • RNA / chemistry
  • Sequence Analysis, RNA / methods

Substances

  • Antibodies
  • RNA
  • DNA