MYC Transactome Mapped by Global Array-based Nuclear Run-on (ANRO - Affymetrix)

We describe and illustrate the use of a method to measure nascent nuclear gene transcription with an Array-based Nuclear Run-On (ANRO) assay using commercial microarray platforms. ANRO measurements in an inducible c-Myc expressing human P493-6 B cell model was performed following 48 hours with or without MYC gene induction. Samples were prepared from either nuclear run-on or total RNA and were applied to Affymetrix Exon arrays and similarly regulated, full length transcripts were demonstrated from each preparation type. Gene expression is compared at a global level after 48 hours with and without tetracycline as measured for both nuclear run-on and total RNA using the Affymetrix Exon microarray platform.