I am looking for antibiotic resistance free plasmid systems and came across this concept of mini-circles. I am wondering what would be the experience of someone who used them, or any practical difficulties?
Any suggestions are welcomed
Thanks
Common Problems with Production of Minicircles
The most common problems with minicircle production are low yield and contamination with parental plasmid or E. coli genomic DNA. These problems can be overcome by slightly modifying specific steps in the protocol. The yield of produced minicircles is inversely proportional to the size of the minicircle particles.
Larger minicircles provide a higher yield, while smaller minicircles give a lower yield. SBI's MC-Easy Minicircle. Production kit is a great way to easily produce your own minicircles.
What were some of the challenges you faced while developing
minicircle technology?
One of the problems we faced was that the arabinose induction used to promote the formation of minicircles never went to completion and at most we could get 80 to 90% minicircle purity without the inclusion of a CsCl based ultracentrifugation step--- a step most laboratory workers like to avoid.
reference: https://www.systembio.com/downloads/SBInsights_vol5.pdf
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