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SRX21541998: GSM7746899: L503-27C-ATAC-rep1; Bigelowiella natans; ATAC-seq
1 ILLUMINA (NextSeq 550) run: 18.9M spots, 1.4G bases, 535.2Mb downloads

External Id: GSM7746899_r1
Submitted by: Genetics, Stanford University
Study: Genome-wide distribution of 5-hydroxymethyluracil and chromatin accessibility in the Breviolum minutum genome
show Abstracthide Abstract
In dinoflagellates, the most unique and divergent nuclear organization among the known diversity of eukaryotes has evolved. The list of highly unusual features of dinoflagellate nuclei and genomes is long -- permanently condensed liquid crystalline chromosomes, in which histones are not the main packaging component, genes organized as very long unidirectional gene arrays, general absence of transcriptional regulation, high abundance of the otherwise very rare DNA modification 5-hydroxymethyluracil (5-hmU), and many others. Most of these fascinating properties were originally identified in the 1970s and 1980s but have received very little attention in recent decades using modern genomic tools. In this work, we address some of the outstanding questions regarding dinoflagellate genome organization by mapping the genome-wide distribution of 5-hmU (using both immunoprecipitation-based and basepair-resolution chemical mapping approaches) and of chromatin accessibility in the genome of the dinoflagellate Breviolum minutum. We find that the 5-hmU modification is preferentially enriched over certain classes of repetitive elements, and also often coincides with the boundaries between gene arrays. It is generally anti-correlated with chromatin accessibility, the levels of which are lower in those regions. We discuss the potential roles of 5-hmU in the functional organization of dinoflagellate genomes and its relationship to the transcriptional landscape of gene arrays. Overall design: ATAC-seq in B. minutum; mapping of 5-hmU in B. minutum using MeDIP-seq and chemical mapping; SMF and ATAC-seq on yeast cells exogenously expressing various dinoflagellate DVNP proteins
Sample: L503-27C-ATAC-rep1
SAMN37205261 • SRS18751397 • All experiments • All runs
Library:
Name: GSM7746899
Instrument: NextSeq 550
Strategy: ATAC-seq
Source: GENOMIC
Selection: other
Layout: PAIRED
Construction protocol: ATAC-seq experiments were performed following the omniATAC protocol. Briefly, ∼100K B. minutum cells were centrifuged at 1,000 g, then resuspended in 500 µL 1× PBS and centrifuged again. Cells were then resuspended in 50 µL ATAC-RSB-Lysis buffer (10 mM Tris-HCl pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% IGEPAL CA-630, 0.1% Tween-20, 0.01% Digitonin) and incubated on ice for 3 minutes. Subsequently 1 mL ATAC-RSB-Wash buffer (10 mM Tris-HCl pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% Tween-20, 0.01% Digitonin) were added, the tubes were inverted several times, and nuclei were centrifuged at 500 g for 5 min at 4◦C. Transposition was carried out by resuspending nuclei in a mix of 25 µL 2× TD buffer (20 mM Tris-HCl pH 7.6, 10 mM MgCl2, 20% Dimethyl Formamide), 2.5 µL transposase (custom produced) and 22.5 µL nuclease-free H2O, and incubating at 37◦C for 30 min in a Thermomixer at 1000 RPM. Transposed DNA was isolated using the MinElute PCR Purification Kit (Qiagen Cat# 28004/28006), and PCR amplified for 10 cycles using the usual ATAC-seq settings. Libraries were purified using the MinElute kit, then sequenced on a Illumina NextSeq 550 instrument as 2x36mers.
Runs: 1 run, 18.9M spots, 1.4G bases, 535.2Mb
Run# of Spots# of BasesSizePublished
SRR2582026018,927,5861.4G535.2Mb2023-10-19

ID:
29025640

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