Isolation of chromatin-associated RNA by sucrose gradient. (A) Flowchart showing a brief summary of the method used for isolation of chromatin RNA. (B) Chromatin isolated from MNase-treated human fibroblast nuclei was run on a 1% agarose gel after sucrose gradient fractionation. The lanes (from left to right) in the agarose gel represent DNA extracted from the chromatin fragments collected from the top to bottom fractions of the sucrose gradient. (C) Chromatin-bound RNA, isolated from the different gradient fractions, was used for semiquantitative RT-PCR with beta-actin primers, and the RT product was loaded according to the gradient fractionation in B; the extreme right lane is a positive control for RT-PCR.
