Preparation of mRNA for expression monitoring.

Autor: Byrne MC; Genetics Institute/Wyeth Research, Cambridge, Massachusetts, USA., Whitley MZ, Follettie MT
Jazyk: angličtina
Zdroj: Current protocols in neuroscience [Curr Protoc Neurosci] 2001 Nov; Vol. Chapter 4, pp. Unit 4.26.
DOI: 10.1002/0471142301.ns0426s16
Abstrakt: The ability to construct comprehensive gene expression profiles comprising hundreds to thousands of genes whose RNA levels are monitored simultaneously represents an exciting new capability in molecular biology. This is accomplished by hybridizing mRNA, which has been quantitatively amplified and labeled with biotin, to DNA chips that display thousands of nucleotides complementary to the mRNAs of interest. In this unit, rationale for starting with poly(A(+)) versus total RNA is discussed, and strategies for choosing oligonucleotides for chip design is presented. Protocols on RNA amplification and labeling, and purifying and quantifying the cDNA and in vitro transcription products are included.
Databáze: MEDLINE