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

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

Blog Article

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 Leg Support Formulas 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 Calendar run-on and total RNA, we propose a semiquantitative PCR approach based on the use of fluorescent primers.