在氯化铯梯度中采用离心步骤对噬菌体λ DNA进行小规模纯化

Marc Mirande, Myriam Lazard, Jean-Pierre Waller
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引用次数: 3

摘要

介绍了一种快速、高效的噬菌体λDNA纯化方法。这种小规模的纯化涉及在氯化铯梯度上分离噬菌体颗粒。使用Airfuge超级离心机,可以在90分钟内轻松完成离心步骤。该方法可对多达12个独立的λDNA进行为期1天的纯化(每个20–40μg)。回收的DNA基本上不含RNA和DNA污染物,可通过限制性内切酶有效切割,并可作为在其他克隆载体中连接DNA片段的起始材料。
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Small-scale purification of bacteriophage λ DNA by an airfuge centrifugation step in cesium chloride gradients

A rapid and efficient procedure for purifying bacteriophage λ DNA is described. This small-scale purification involves isolation of bacteriophage particles on cesium chloride gradients. Using an Airfuge ultracentrifuge, the centrifugation step can be readily achieved in 90 minutes. The method allows a 1-day purification of up to 12 independent λ DNA (20–40 μg each). The recovered DNA, essentially devoid of RNA and DNA contaminants, is efficiently cut by restriction endonucleases and can serve as starting material for the ligation of DNA fragments in other cloning vehicles.

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Gene targeting in murine embryonic stem cells: Introduction of specific alterations into the mammalian genome A solution hybridization method for quantification of mRNAs: Determining the amount and stability of oncogene mRNA The use of transgenic mice for short-term, in vivo mutagenicity testing Author index volume 7 Subject index volume 7
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