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Updated: Jul 20, 2026

Production of Double-stranded DNA Ministrings
Published on: February 29, 2016
Protocol for minicircle production for gene therapy without subsequent cleanup steps
Jonathan Do1, Gautam Verma1, Yasmin Maurice1
1Phoenix Children's Research Institute, University of Arizona College of Medicine-Phoenix, Phoenix, AZ, USA.
Abstract:
Current techniques for producing minicircles have low yields because of the cleanup steps required. Here, we present a protocol that can increase minicircle yield up to 10-fold by eliminating post-induction cleanup steps. We describe steps for bacterial culture, same-day arabinose induction, and DNA harvesting. This protocol is relevant for all minicircles produced from parental plasmids containing attB, attP, and 32 copies of Isce-I endonuclease sites in the minicircle-producing bacteria, ZYCY10PS3T2.
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