Bio-Protocol (Oct 2016)

PCR-based Assay for Genome Integrity after Methyl Methanesulfonate Damage in Physcomitrella patens

  • Masaki Odahara,
  • Takayuki Inouye,
  • Yoshiki Nishimura,
  • Yasuhiko Sekine

DOI
https://doi.org/10.21769/BioProtoc.1954
Journal volume & issue
Vol. 6, no. 19

Abstract

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In plant cells, genomic DNA exists in three organelles: the nucleus, chloroplast, and mitochondrion. Genomic DNA can be damaged by endogenous and exogenous factors, but the damaged DNA can be repaired by DNA repair systems. To quantify the extent of their repair activity of on individual genomic DNA, a PCR-based assay utilizing long amplicons is valuable for evaluable. This assay is based on the inhibitory effects of methyl methanesulfonate (MMS)-induced DNA damage on the amplicons. This assay is useful for assessing DNA double-strand repair pathways, such as homologous recombination repair, as it detects DNA double-strand breaks produced by MMS in vivo.