Strand-seq of human embryonic stem cells and differentiated cells in ATRX wt and ATRX ko cells

ABSTRACT: Briefly, human embryonic stem cells (hESCs) were collected after BrdU pulse and resuspended in nuclei staining buffer (100 mM Tris-HCl pH 7.4, 150 mM NaCl, 1 mM CaCl2, 0.5 mM MgCl2, 0.1% NP-40, and 2% bovine serum albumin [Sigma-Aldrich] supplemented with 10 μg/mL Hoechst 33258 [Enzo] and 10 μg/mL propidium iodide [Sigma-Aldrich]. Single nuclei were sorted into 5 μL ProFreeze-CDM freeze medium [Lonza] + 7.5% dimethyl sulfoxide in 96-well skirted PCR plates [4titude], based on low propidium iodide (G1 phase) and low Hoechst (BrdU-induced quenching) fluorescence using an Influx cell sorter [BD Biosciences]. Strand-seq libraries were prepared using an Agilent Bravo liquid handling platform as described before (Sanders et al., 2017). For each experiment, 96 libraries were pooled and 250-450 bp

INSTRUMENT(S): Illumina HiSeq 2500

ORGANISM(S): Homo sapiens

SUBMITTER: Antonio Maffia 

PROVIDER: E-MTAB-12582 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress