Rescue of S/MAR-containing nonviral episomal expression vectors

Cold Spring Harb Protoc. 2012 Jun 1;2012(6):709-11. doi: 10.1101/pdb.prot069518.

Abstract

The episomal status of S/MAR (scaffold/matrix attached region)-based vectors can be confirmed by several methods including Southern blots, fluorescence in situ hybridization (FISH) analysis, or plasmid rescue experiments. In rescue experiments, genomic DNA (gDNA) or DNA from Hirt extracts is isolated from cell clones or mixed populations in which S/MAR plasmids are stably established. Bacteria are transformed with this DNA and if episomal plasmid DNA (pDNA) is present, resistant bacterial colonies will form.

MeSH terms

  • Escherichia coli / genetics
  • Genetic Engineering / methods*
  • Genetic Vectors / analysis*
  • Genetic Vectors / isolation & purification*
  • Plasmids / analysis*
  • Plasmids / isolation & purification*
  • Transformation, Bacterial*
  • Transformation, Genetic*