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通过微核选择性捕获无着丝粒片段为纯化染色体外扩增DNA提供了一种快速方法。

Selective capture of acentric fragments by micronuclei provides a rapid method for purifying extrachromosomally amplified DNA.

作者信息

Shimizu N, Kanda T, Wahl G M

机构信息

Gene Expression Laboratory, Salk Institute for Biological Studies, La Jolla, California 92037, USA.

出版信息

Nat Genet. 1996 Jan;12(1):65-71. doi: 10.1038/ng0196-65.

Abstract

The amplification and overexpression of a number of oncogenes is strongly associated with the progression of a variety of different cancers. We now present a strategy to purify amplified DNA on double minute chromosomes (DMs) to enable analysis of their prevalence and contribution to tumourigenesis. Using cell lines derived from four different tumour types, we have developed a general and rapid method to purify micronuclei that are known to entrap extrachromosomal elements. The isolated DNA is highly enriched in DM sequences and can be used to prepare probes to localize the progenitor single copy chromosomal regions. The capture of DMs by micronuclei appears to be dependent on their lack of a centromere rather than their small size.

摘要

许多癌基因的扩增和过表达与多种不同癌症的进展密切相关。我们现在提出一种策略,用于纯化双微体染色体(DMs)上的扩增DNA,以便分析其在肿瘤发生中的普遍性和作用。利用源自四种不同肿瘤类型的细胞系,我们开发了一种通用且快速的方法来纯化已知会捕获染色体外元件的微核。分离出的DNA高度富集DM序列,可用于制备探针以定位祖代单拷贝染色体区域。微核对DMs的捕获似乎取决于它们缺乏着丝粒而非其小尺寸。

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