Rescue of S/MAR-Containing Nonviral Episomal Expression Vectors
- 1Centre for Biomedical Education and Research, Institute of Cell Biology, University of Witten/Herdecke, 58453 Witten, Germany
- 2Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, 85764 Oberschleißheim, Germany
- 3Helios Medical Centre Wuppertal, Paediatrics Centre, 42117 Wuppertal, Germany
- 4Institute for Virology and Microbiology, ZBAF, University of Witten/Herdecke, 58453 Witten, Germany
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.
Footnotes
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↵5 Correspondence: claudia.hagedorn{at}uni-wh.de
- © 2012 Cold Spring Harbor Laboratory Press
