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Sample GSM1195398 Query DataSets for GSM1195398
Status Public on Jul 26, 2013
Title seq-SDQ3166_LIN37_N2_LTemb_ChIP_Rep2
Sample type SRA
 
Source name seq-SDQ3166_LIN37_N2_LTemb_ChIP_Rep2
Organism Caenorhabditis elegans
Characteristics strain: N2
developmental stage: Late Embryos
genotype: wild type
Sex: Hermaphrodite
Growth protocol Worm_embryo_growth_and_harvest_vSS7. Embryos were prepared by bleaching from gravid N2 adults grown in standard S-basal media liquid culture. Embryos were incubated for 3.5 hours in M9 then frozen in liquid nitrogen.
Extracted molecule genomic DNA
Extraction protocol Worms are frozen, ground, and crosslinked for 10 minutes in 1% formaldehyde. Later,washed pellets are resuspended in FA buffer and subjected to sonication in a DiagenodeBioruptor (40 pulses of 30 seconds with 1 minute rests in between at full power). Extractsare then spun down and the soluble fraction is stored for quality tests and future ChIP.
Worm_chromatin_immunoprecipitation_vSS4. 0.3-3 mg extract + 1% sarkosyl was used for each ChIP with 10% taken as input directly into elution buffer (1% SDS in TE, 250 mM NaCl). Antibody was added to each IP sample and incubated overnight at 4ºC. Immune complexes were incubated (2 hrs at 4ºC) with 50 ?l of IgG dynabeads (Dyna), and washed 5 minutes with 1.5 mL of each of the following solutions: ChIP Buffer, ChIP Buffer + 1 M NaCl, ChIP Buffer + 500 mM NaCl, TEL Buffer (10 mM Tris-HCl pH 8.0, 250 mM LiCl, 1% NP-40, 1% sodium deoxycholate, 1 mM EDTA), and 2X TE (10 mM Tris-HCl pH 8.0, 1 mM EDTA). Samples were eluted twice with 150 ?L elution buffer for 15 minutes at 65ºC. Samples were treated with RNAse for 30 minutes at 37ºC. Samples were treated with proteinase K at 55ºC for 1-2 hrs then transfer to 65ºC overnight to reverse crosslinks. DNA was cleaned up using Zymo kit. For a detailed protocol see http://www.modencode.org/.
Short description for UNC NimbleGen Database:Both ChIP and input DNA samples have their ends blunted and phosphorylated with KlenowDNAP, T4 DNAP, and T4 PNK. 3?-dA overhangs are then added with Klenow Fragment (3? to 5?exo minus), and Illumina adapters are subsequently ligated to the ends. Next, PCRamplification is performed using Illumina 1.1 and 2.1 primers, for a total of 18 cycles. AmplifiedDNA libraries are size-selected on a 2% agarose gel so that fragments sized between 250-350bp are obtained; gel extraction is carried out at room temperature.
 
Library strategy ChIP-Seq
Library source genomic
Library selection ChIP
Instrument model Illumina Genome Analyzer
 
Description ChIP DNA; Antibody information listed below: official name: BH00003 LIN37;target name: LIN-37;host: Rabbit;antigen: Full length;clonal: Polyclonal;purified: Affinity;company: Other;catalog: Bob Horvitz;input DNA
Data processing to be updated
 
Submission date Jul 25, 2013
Last update date May 15, 2019
Contact name DCC modENCODE
E-mail(s) [email protected]
Phone 416-673-8579
Organization name Ontario Institute for Cancer Research
Lab modENCODE DCC
Street address MaRS Centre, South Tower, 101 College Street, Suite 800
City Toronto
State/province Ontario
ZIP/Postal code M5G 0A3
Country Canada
 
Platform ID GPL9309
Series (1)
GSE49204 seq-SDQ3166_LIN37_N2_LTemb
Relations
Reanalyzed by GSE95071
BioSample SAMN02264731
SRA SRX529221

Supplementary file Size Download File type/resource
GSM1195398_seq-SDQ3166_LIN37_N2_LTemb_2_peaks.gff.gz 28.1 Kb (ftp)(http) GFF
GSM1195398_seq-SDQ3166_LIN37_N2_LTemb_2_treat_afterfiting_all.wig.gz 29.7 Mb (ftp)(http) WIG
SRA Run SelectorHelp
Raw data are available in SRA
Processed data provided as supplementary file
Processed data are available on Series record

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