Purification of BAC DNA for high-efficiency transgenesis
Purification of BAC DNA for high-efficiency transgenesis
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DOI：
10.2144/000113763
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发表时间：
2011-11-01
期刊：
影响因子：
2.7
通讯作者：
Bhat, Suraj P.
中科院分区：
文献类型：
--
作者：
Gangalum, Rajendra K.;Jing, Zhe;Bhat, Suraj P.
细菌人工染色体（BAG）转基因的一个尚未解决的瓶颈是由于操作的BAG DNA质量不佳而导致创始小鼠的低效率产生。使用微型凝胶电泳和电洗脱，绕过CsCl（2）离心，柱层析和树脂纯化，我们使用了RECOCHIP，一种市售的透析盒用于BAG DNA的纯化，产生转基因创始人高达80%的效率。
An unresolved bottleneck in bacterial artificial chromosome (BAG) transgenesis is low efficiency generation of founder mice because of suboptimal quality of the manipulated BAG DNA. Using mini-gel electrophoresis and electro-elution that circumvents CsCl(2) centrifugation, column chromatography, and resin purifications, we have used RECOCHIP, a commercially available dialysis cassette for the purification of BAG DNA that generates transgenic founders with up to 80% efficiency.