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Status |
Public on Aug 05, 2014 |
Title |
Genotyping of H1. UW, Stamlab (NREMC). H1 cell line; JS-ESCH1-genotype |
Sample type |
genomic |
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Source name |
Human embryonic stem cells received from the James Thompson laboratory. Cells are from the H1 line.; JS-ESCH1-genotype
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Organism |
Homo sapiens |
Characteristics |
disease: None biomaterial_provider: James Thompson Laboratory biomaterial_type: Cell Line line: H1 lineage: Embryonic Stem Cell differentiation_stage: None differentiation_method: None passage: Between 20 and 40 medium: mTeSER Sex: Male batch: H1U
|
Treatment protocol |
None
|
Growth protocol |
http://roadmapepigenomics.org/protocols
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Extracted molecule |
genomic DNA |
Extraction protocol |
Qiagen DNeasy Blood and Tissue kit (cat #69506).
|
Label |
Illumina protocol
|
Label protocol |
The chip underwent staining and extension in capillary flow-through chambers. Allele-specific single-base extension of the oligos on the beadchip, using the captured DNA as template, incorporated detectable labels on the beadchip and determined the genotype call for the sample.
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Hybridization protocol |
200ng of genomic DNA was first denatured and subsequently neutralized to prepare it for amplification. The denatured DNA was then isothermally amplified in an overnight step at 37°C. Next, amplified DNA was enzymatically fragmented for 60 minutes at 37°C using an end-point fragmentation process. The fragmented DNA was then precipitated with isopropanol, allowed to air dry, then resuspended in hybridization buffer. Eight samples were subsequently applied to each HumanOmni2.5-8v1.1 beadchip, which were kept separate with an IntelliHyb seal. The prepared beadchip was incubated overnight in a hybridization oven at 48°C for 16-24 hours with rocking. The amplified and fragmented DNA samples annealed to locus-specific 50mers during hybridization. Following hybridization, unhybridized and non-specifically hybridized DNA was washed away, and the chip was prepared for staining and extension.
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Scan protocol |
Beadchips were scanned using an Illumina iScan+ with ICS v3.2.28
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Description |
sample_term_id: EFO_0003042 assay_term_id: OBI_0001393 nucleic_acid_term_id: SO_0000352 Genotyping EDACC Genboree Experiment Page: http://genboree.org/java-bin/project.jsp?projectName=XML%20Submissions%2FUniversity%20of%20Washington%2FEXPERIMENT%2FEDACC.19800 sample alias: H1-13 EDACC Genboree Sample Page: http://genboree.org/java-bin/project.jsp?projectName=XML%20Submissions%2FUCSD%2FSAMPLE%2FEDACC.3425
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Data processing |
Intensity data was extracted with Illumina GenomeStudio software (GenomeStudio v2011.1 with Genotyping Module v1.9.4) with a GenCall cutoff of 0.15.
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Submission date |
Jul 31, 2014 |
Last update date |
Jan 29, 2015 |
Contact name |
Northwest REMC |
E-mail(s) |
rharris1@bcm.tmc.edu
|
Organization name |
University of Washington
|
Street address |
-
|
City |
Seattle |
State/province |
WA |
ZIP/Postal code |
98195 |
Country |
USA |
|
|
Platform ID |
GPL18952 |
Series (1) |
GSE18927 |
University of Washington Human Reference Epigenome Mapping Project |
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Relations |
BioSample |
SAMN00114950 |