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Status |
Public on Aug 13, 2020 |
Title |
HTZ1.lin35.rep2 |
Sample type |
SRA |
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Source name |
HTZ1.lin35.whole worm
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Organism |
Caenorhabditis elegans |
Characteristics |
strain: lin-35 (n745) developmental stage: starved L1 growth temperature: 25°C tissue: whole worm chip antibody: HTZ1 sample id: CG053
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Growth protocol |
Starved L1 animals were collected by bleaching synchronized adults grown in liquid culture using standard S-basal medium with HB101 E. coli at 20°C. Following bleaching, eggs were hatched at 25°C in M9 and starved L1s were sucrose floated and collected 20-22 hours following hatching by flash freezing in liquid nitrogen.
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Extracted molecule |
genomic DNA |
Extraction protocol |
Frozen starved L1 worms were ground to a powder, which was incubated in 1.5 mM EGS (Pierce 21565) in PBS for 8 minutes, followed by the addition of formaldehyde to a final concentration of 1%, and incubated for a further 8 minutes. The fixation was quenched for 5 minutes by the addition of 0.125 M glycine. Fixed tissue was washed 2X with PBS with protease inhibitors (Roche EDTA-free protease inhibitor cocktail tablets 05056489001) and once in FA buffer (50 mM Hepes pH7.5, 1 mM EDTA, 1% TritonX-100, 0.1% sodium deoxycholate, and 150 mM NaCl) with protease inhibitors (FA+), then resuspended in 1 ml FA+ buffer per 1 ml of ground worm powder. The extract was sonicated to an average size of ~250 base pairs using a Bioruptor Pico (Diagenode), and 10-20 ug of DNA was used per ChIP reaction. ChIP DNA is blunt ended, A-tailed, ligated to adaptors, amplified by PCR, then size selected using AMPure beads. Illumina TruSeq adaptors and barcodes were used.
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Library strategy |
ChIP-Seq |
Library source |
genomic |
Library selection |
ChIP |
Instrument model |
Illumina HiSeq 1500 |
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Description |
HTZ1.lin35.rep2.CG053
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Data processing |
ChIP-seq reads were aligned to a concatenated ce11+cb3 assembly of the C. elegans and C. briggsae genomes using BWA v. 0.7.7 with default settings (BWA-backtrack algorithm) (Li and Durbin, 2010). The SAMtools v. 0.1.19 ‘view’ utility (Li et al., 2009) was used to convert the alignments to BAM format. Normalized mapq10 ChIP-seq coverage tracks were generated using the BEADS algorithm (Cheung et al., 2011). Genome_build: ce11 + cb3 (concatenated) Supplementary_files_format_and_content: BEADS-normalized genome-wide linear read coverage (from reads with MAPQ value >= 10)
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Submission date |
Jul 27, 2020 |
Last update date |
Aug 13, 2020 |
Contact name |
Francesco Nicola Carelli |
E-mail(s) |
[email protected]
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Organization name |
University of Cambridge
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Lab |
Julie Ahringer
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Street address |
Henry Wellcome Building of Cancer and Developmental Biology, Tennis Court Road
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City |
Cambridge |
ZIP/Postal code |
CB2 1QN |
Country |
United Kingdom |
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Platform ID |
GPL18730 |
Series (2) |
GSE155189 |
DREAM represses distinct targets by cooperating with different THAP domain proteins [CHIP-seq] |
GSE155191 |
DREAM represses distinct targets by cooperating with different THAP domain proteins |
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Relations |
BioSample |
SAMN15651115 |
SRA |
SRX8832066 |
Supplementary file |
Size |
Download |
File type/resource |
GSM4697037_HTZ1.lin35.rep2.CG053.BEADSQ10NU_linear.bw |
235.9 Mb |
(ftp)(http) |
BW |
SRA Run Selector |
Raw data are available in SRA |
Processed data provided as supplementary file |
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