RNA提取、cDNA制备及CFTR转录本分析方法。

Methods for RNA extraction, cDNA preparation and analysis of CFTR transcripts.

作者信息

Ramalho Anabela S, Beck Sebastian, Farinha Carlos M, Clarke Luka A, Heda Ghanshyam D, Steiner Bernhard, Sanz Javier, Gallati Sabina, Amaral Margarida D, Harris Ann, Tzetis Maria

机构信息

Centre of Human Genetics, National Institute of Health, Lisboa, Portugal.

出版信息

J Cyst Fibros. 2004 Aug;3 Suppl 2:11-5. doi: 10.1016/j.jcf.2004.05.004.

DOI:10.1016/j.jcf.2004.05.004

PMID:15463918

Abstract

The scope of this article is to outline some of the basic methods for good quality RNA preparation from mammalian tissues and cells (including epithelial cells). Additionally, we give an outline of common techniques of measuring CFTR gene expression such as quantitative and semi-quantitative reverse transcription (RT) PCR and ribonuclease protection assay (RPA). These methods are designed to detect low abundance transcripts, which apply to CFTR mRNA in most cell types and tissues.

摘要

本文的范围是概述从哺乳动物组织和细胞（包括上皮细胞）中制备高质量RNA的一些基本方法。此外，我们还概述了测量CFTR基因表达的常用技术，如定量和半定量逆转录（RT）PCR以及核糖核酸酶保护分析（RPA）。这些方法旨在检测低丰度转录本，适用于大多数细胞类型和组织中的CFTR mRNA。