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通过激光介导的单细胞操作鉴定表达基因。

Identification of expressed genes by laser-mediated manipulation of single cells.

作者信息

Schütze K, Lahr G

机构信息

Laser Laboratory and Molecular Biology, First Medical Department, Academic Hospital München-Harlaching, Germany.

出版信息

Nat Biotechnol. 1998 Aug;16(8):737-42. doi: 10.1038/nbt0898-737.

Abstract

We describe a rapid noncontact method for the capture of single cells or small tissue areas of any size or shape directly within the cap of a common microfuge tube. Prior to the laser-mediated transfer, the specimen is isolated by laser microbeam microdissection, forming a clear-cut gap around the selected area. Laser treatment does not impair subsequent RNA analysis. We have used this method to isolate a single cell from archival colon adenocarcinoma, and were able to detect point mutations within codon 12 of c-Ki-ras2 mRNA after nested RT-PCR analysis.

摘要

我们描述了一种快速非接触方法,可直接在普通微量离心管的管帽内捕获任何大小或形状的单个细胞或小组织区域。在激光介导转移之前,通过激光微束显微切割分离标本,在选定区域周围形成清晰的间隙。激光处理不会损害后续的RNA分析。我们已使用此方法从存档的结肠腺癌中分离出单个细胞,并在巢式RT-PCR分析后能够检测到c-Ki-ras2 mRNA第12密码子内的点突变。

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