DNase I SIM: A Simplified In-Nucleus Method for DNase I Hypersensitive Site Sequencing

TitleDNase I SIM: A Simplified In-Nucleus Method for DNase I Hypersensitive Site Sequencing
Publication TypeBook Chapter
Year of Publication2017
AuthorsFilichkin, SA, Megraw, M
Book TitleMethods Mol Biol
Volume1629
Pagination141-154
ISBN Number1940-6029 (Electronic)1064-3745 (Linking)
Abstract

Identifying cis-regulatory elements is critical in understanding the direct and indirect interactions that occur within gene regulatory networks. Current approaches include DNase-seq, a technique that combines sensitivity to the nonspecific endonuclease DNase I with high-throughput sequencing to identify regions of regulatory DNA on a genome-wide scale. Yet, challenges still remain in processing recalcitrant tissues that have low DNA content. Here, we describe DNase I SIM (for Simplified In-nucleus Method), a protocol that simplifies and facilitates generation of DNase-seq libraries from plant tissues for high-resolution mapping of DNase I hypersensitive sites. By removing steps requiring the use of gel agarose plugs in DNase-seq, DNase I SIM reduces the time required to perform the protocol by at least 2 days, while also making possible the processing of difficult plant tissues including plant roots.

Short TitleMethods in molecular biology (Clifton, N.J.)