Sandeep Raha , Frank Merante , Gerald Proteau , Juta K. Reed
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引用次数: 35
摘要
描述了从培养细胞中分离完整的细胞总RNA的快速程序。该方法结合了同时破坏细胞和提取核酸的一个步骤,使用苯酚和含有100 mM LiCl的缓冲液。总细胞RNA可在约2小时内分离。RNA的产量和质量可与更广泛使用的方法相媲美,这些方法需要大量的准备步骤,如使用硫氰酸胍提取和随后的CsCl梯度离心。使用我们的方法分离的RNA包含长达10 kb的转录本,适合于Northern分析。这个过程也产生高分子量的DNA,这是一个合适的底物限制内切酶。
Simultaneous isolation of total cellular RNA and DNA from tissue culture cells using phenol and lithium chloride
A rapid procedure for the isolation of intact total cellular RNA from cultured cells is described. This method combines the simultaneous disruption of cells and extraction of nucleic acids in a single step with the use of phenol and a buffer containing 100 mM LiCl. Total cellular RNA can be isolated in approximately 2 hours. The yield and quality of the RNA is comparable to the more widely employed methods requiring extensive preparatory steps such as extraction using guanidinium thiocyanate and subsequent CsCl gradient centrifugation. The RNA isolated using our procedure contains transcripts up to 10 kb in length and is suitable for Northern analysis. This procedure also yields high-molecular-weight DNA, which is a suitable substrate for restriction endonucleases.