文章基本信息

标题：Detection of DNA point mutations and mRNA expression levels by rolling circle amplification in individual cells
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作者：Allen T. Christian ; Melissa S. Pattee ; Christina M. Attix 等
期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：2001
卷号：98
期号：25
页码：14238-14243
DOI：10.1073/pnas.251383598
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：Rolling circle amplification has been useful for detecting point mutations in isolated nucleic acids, but its application in cytological preparations has been problematic. By pretreating cells with a combination of restriction enzymes and exonucleases, we demonstrate that rolling circle amplification in situ can detect gene copy number and single base mutations in fixed cells with efficiencies up to 90%. It can also detect and quantify transcribed RNA in individual cells, making it a versatile tool for cell-based assays.