title: chromatin associated RNAs
design: chromatin associated RNAs
library_construction_protocol: Chromatin assembly and accessibility analysis were performed as described elsewhere (Becker and Wu, 1992; Rhodes and Laskey, 1989; Stewart et al., 1991) For RNA isolation, chromatin was fractionated in a linear sucrose gradient (15-40%; 12 h; 7500g; 4�C). Fractions containing the chromatin associated RNA were de-proteinized and treated with DNase I. Isolated RNA was enriched for fragments < 200 bp and strand-specific cDNA libraries were prepared using the ExactStart Small RNA Cloning Kit (Epicentre). Libraries included small coding as well as non-coding RNAs and fragments of longer primary transcripts. The cDNA was amplified with hybrid primers containing sequences required for the Illumina platform. PCR products were cleaned up with AMPure XP beads (Beckman Coulter).
attributes.tag: Experimental Factor: RNA
attributes.value: total embryonal RNA