GSM1518776: n2a-Dali-R1; Mus musculus; OTHER
CHART Enrichment and RNase H Mapping experiments were performed as previously described (Simon, 2013, Capture hybridization analysis of RNA targets (CHART). Curr Protoc Mol Biol Chapter 21: Unit 21 25). CHART extract was prepared from approximately 3 × 10^8  N2A cells per pull down and hybridized with 810 pmol biotinylated oligonucleotide cocktail overnight with rotation at room temperature. Complexes were captured using 250ml MyOneC1 streptavidin beads (Invitrogen) overnight at room temperature with rotation. Bound material was extensively washed and eluted using RNase H (New England Biolabs) for 30 min at room temperature. Samples were treated with Proteinase K and cross-links were reversed. DNA was sheared to an average fragment size of 150-300 bp using a Bioruptor (Diagenode). Fragmented DNA was end-repaired, A-tailed and adapter-ligated before size selection and amplification. The prepared libraries were QC’ed and multiplexed before paired end sequencing over one lane of a Illumina HiSeq flow cell. For more information contact the High-Throughput Genomics Group at the Wellcome Trust Centre for Human Genetics, Oxford.
GEO
GSM1518776
GEO Accession
GSM1518776