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.
Protocol for minicircle production for gene therapy without subsequent cleanup steps.
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作者：Do Jonathan, Verma Gautam, Maurice Yasmin, Abdumanobova Muniba, Deng Zicheng, Kalin Tanya V, Kalinichenko Vladimir V
| 期刊： | STAR Protocols | 影响因子： | 1.300 |
| 时间： | 2025 | 起止号： | 2025 Jul 24; 6(3):103982 |
| doi： | 10.1016/j.xpro.2025.103982 | ||
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