Article Title: Chromatin run-on reveals the transcriptional etiology of glioblastoma multiforme
Figure Lengend Snippet: ChRO-seq and leChRO-seq measure primary transcription in isolated chromatin. (a) Isolated chromatin is incubated with biotinylated rNTPs, purified by streptavidin beads, and sequenced from the 3’ end. leChRO-seq degrades existing RNA, extends nascent transcripts an average of 100 bp, and sequences RNAs from the 5’ end. (b and c) Comparison between matched ChRO-seq and PRO-seq in annotated gene bodies (b) or at the peak of paused Pol II (c) in units of reads per kilobase per million mapped. (d) Comparison between ChRO-seq (top three tracks), PRO-seq (center), and H3K27ac ChlP-seq, DNase-I-seq, and RNA-seq (bottom). dREG-HD shows the raw signal fordREG (gray) and imputed DNase-I hypersensitivity signal (dark red). (e) The distribution of read lengths from ChRO-seq (blue) and leChRO-seq (pink) in a 30 year old primary GBM.
Article Snippet: Nascent RNA was purified by binding streptavidin beads (NEB # S1421S) and washed as described .
Techniques: Isolation, Incubation, Purification, RNA Sequencing Assay