PDF(Pubmed)
Abstract：
We recently identified a class of small cytosolic double-stranded DNA (scDNA) approximately 20-40 bp in size in human and mouse cells. Here, we present a protocol for scDNA isolation from cultured murine cells. We describe steps for cytosolic compartment separation, DNA isolation in the cytosolic fraction using phenol-chloroform extraction, and ethanol precipitation. We then detail procedures for denaturing purified cytosolic DNA through urea polyacrylamide gel electrophoresis and obtaining scDNA in the cytosolic DNA fraction via gel purification. For complete details on the use and execution of this protocol, please refer to Liu et al.1.
摘要：
我们最近在人和小鼠细胞中鉴定了一类大小约20-40bp的小胞质双链DNA(scDNA)。这里,我们提出了从培养的鼠细胞中分离scDNA的方案。我们描述了胞质区室分离的步骤，使用苯酚-氯仿提取在胞质部分中分离DNA，和乙醇沉淀。然后，我们详细介绍了通过尿素聚丙烯酰胺凝胶电泳使纯化的细胞溶质DNA变性并通过凝胶纯化在细胞溶质DNA部分中获得scDNA的程序。有关此协议的使用和执行的完整详细信息，请参考刘等人。1。
