Format

Send to

Choose Destination
Biotechniques. 2000 Nov;29(5):1012-4, 1016-7.

Nuclear run-on assay using biotin labeling, magnetic bead capture and analysis by fluorescence-based RT-PCR.

Author information

1
Laboratorio di Genetica Molecolare, Istituto G. Gaslini, Università di Genova, Italy. aldamar@tin.it

Abstract

In this report, we present a fluorescence-based approach to the assessment of cellular gene expression and transcription rates. Nuclear run-on was performed by supplying biotin-16-UTP to nuclei, and labeled transcripts were bound to streptavidin-coated magnetic beads. Total cDNA was then synthesized by means of random hexamer primed reverse transcription of captured molecules. To monitor transcript abundance in cDNA, both from nuclear run-on and total RNA, we propose a semiquantitative PCR approach based on the use of fluorescent primers.

PMID:
11084863
DOI:
10.2144/00295st02
[Indexed for MEDLINE]
Free full text

Supplemental Content

Full text links

Icon for Atypon
Loading ...
Support Center