Nuclear Run-On Assay Using Biotin Labeling, Magnetic Bead Capture and Analysis by Fluorescence-Based RT-PCR | Zendy

Giovanna Patrone | Zendy; Francesca Puppo | Zendy; Roberto Cusano | Zendy; Monica Scaranari | Zendy; Isabella Ceccherini | Zendy; Aldamaria Puliti | Zendy; Roberto Ravazzolo | Zendy

AI Assistant Blog Pricing

Home ZAIA Blog

Open Access

Nuclear Run-On Assay Using Biotin Labeling, Magnetic Bead Capture and Analysis by Fluorescence-Based RT-PCR

Author(s) -

Giovanna Patrone,

Francesca Puppo,

Roberto Cusano,

Monica Scaranari,

Isabella Ceccherini,

Aldamaria Puliti,

Roberto Ravazzolo

Publication year - 2000

Publication title -

biotechniques

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.617

H-Index - 131

eISSN - 1940-9818

pISSN - 0736-6205

DOI - 10.2144/00295st02

Subject(s) - biotin , complementary dna , microbiology and biotechnology , random hexamer , fluorescence , streptavidin , reverse transcription polymerase chain reaction , reverse transcriptase , biology , messenger rna , real time polymerase chain reaction , biotinylation , rna , chemistry , gene , genetics , physics , quantum mechanics

In this report, we present a fluorescencebased 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.

The content you want is available to Zendy users.

Already have an account? Click here to sign in.

Having issues? You can contact us here

Accelerating Research