Rescue of S/MAR-containing nonviral episomal expression vectors.
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.
Centre for Biomedical Education and Research, Institute of Cell Biology, University of Witten/Herdecke, 58453 Witten, Germany.
Claudia
C
Hagedorn
Armin
A
Baiker
Jan
J
Postberg
Anja
A
Ehrhardt
Hans J
HJ
Lipps
D004926:Escherichia coli / Q000235:genetics; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000032:analysis* / Q000302:isolation & purification*; D010957:Plasmids / Q000032:analysis* / Q000302:isolation & purification*; D014169:Transformation, Bacterial; D014170:Transformation, Genetic