一种使用均一化cDNA文库的大规模原位杂交系统。

A large-scale in situ hybridization system using an equalized cDNA library.

作者信息

Komiya T, Tanigawa Y, Hirohashi S

机构信息

ERATO, Japan Science and Technology Corporation (JST), 5-9-4 Tokodai, Tsukuba, 300-26, Japan.

出版信息

Anal Biochem. 1997 Dec 1;254(1):23-30. doi: 10.1006/abio.1997.2399.

DOI:10.1006/abio.1997.2399

PMID:9398341

Abstract

We have developed a large-scale in situ hybridization system in which all the procedures are carried out on a 96-well format: digoxigenin-labeled probes were synthesized from PCR-amplified templates, sections were mounted on 96-well plates, and hybridization and immunohistochemistry protocols were carried out in each well of the plates. This system in combination with equalized (normalized) cDNA library as the source for the probes enables us to identify the cellular distribution of many mRNAs in various tissues rapidly and efficiently. Thus, this system may be a novel cloning method of identify genes differentially expressed in many tissues. In addition, this system has a potential to be automated.

摘要

我们开发了一种大规模原位杂交系统，其中所有程序均以96孔板形式进行：从PCR扩增模板合成地高辛标记探针，将切片置于96孔板上，并在板的每个孔中进行杂交和免疫组织化学实验方案。该系统与作为探针来源的均衡（标准化）cDNA文库相结合，使我们能够快速有效地鉴定多种组织中许多mRNA的细胞分布。因此，该系统可能是一种鉴定在多种组织中差异表达基因的新型克隆方法。此外，该系统具有自动化的潜力。