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| Status |
Public on Oct 07, 2010 |
| Title |
Whole genome shotgun bisulfite sequencing of the H1 cell line treated with BMP4 for 5 days; methylC-seq_h1+bmp4_r2a |
| Sample type |
SRA |
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| Source name |
H1 Cell Line treated with BMP4, biological replicate 2 (r2); methylC-seq_h1+bmp4_r2a
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| Organism |
Homo sapiens |
| Characteristics |
molecule: genomic DNA
disease: None
biomaterial_provider: Thomson Laboratory
biomaterial_type: Cell Line
line: H1
lineage: NA
differentiation_stage: embryonic stem cell differentiated by treatment with BMP4
differentiation_method: H1 cells were cultured in TeSR containing 50 ng/ml BMP4 for 5 days
passage: 25, including 10 passages in TeSR
medium: TeSR
Sex: Male
batch: Replicate 2
experiment_type: DNA Methylation
extraction_protocol: Qiagen DNeasy mini kit, performed as per manufacturer's instructions
extraction_protocol_type_of_sonicator: Covaris S2
extraction_protocol_sonication_cycles: Standard fragment express, 6 cycles
dna_preparation_initial_dna_qnty: 5 µg
dna_preparation_fragment_size_range: 100-150
dna_preparation_adaptor_sequence: A: 5' P-GATCGGAAGAGCTCGTATGCCGTCTTCTGCTTG, B: 5' ACACTCTTTCCCTACACGACGCTCTTCCGATCT
dna_preparation_adaptor_ligation_protocol: 16ËšC for 16 hours with T4 DNA ligase (New England Biolabs)
dna_preparation_post-ligation_fragment_size_selection: Two rounds of purification with AMPure XP beads (Agencourt)
bisulfite_conversion_protocol: Invitrogen MethylCode
bisulfite_conversion_percent: 99.5% of cytosines converted based on shotgun sequencing of unmethylated lambda phage control spiked into original genomic DNA sample
library_generation_pcr_template_conc: >One third of the adapter-ligated, bisulfite converted DNA was used in each of three 50 µl PCR reaction
library_generation_pcr_polymerase_type: Stratagene Pfu Turbo Cx
library_generation_pcr_thermocycling_program: 95ËšC 2 min; 98ËšC 30 sec, 4 cycles of 98ËšC 15 sec, 60ËšC 30 sec, 72ËšC 4 min; 72ËšC 10 min
library_generation_pcr_number_cycles: 4
library_generation_pcr_f_primer_sequence: 5' AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCT
library_generation_pcr_r_primer_sequence: 5' CAAGCAGAAGACGGCATACGAGCTCTTCCGATCT
library_generation_pcr_primer_conc: 25 µM
library_generation_pcr_product_isolation_protocol: Two rounds of purification with AMPure XP beads (Agencourt)
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| Extracted molecule |
genomic DNA |
| Extraction protocol |
Library construction protocol: Five µg of genomic DNA was extracted from frozen cell pellets using the DNeasy Mini Kit (Qiagen, Valencia, CA) and spiked with 25 ng unmethylated cl857 Sam7 Lambda DNA (Promega, Madison, WI). The DNA was fragmented with a Covaris S2 (Covaris, Woburn, MA) to 100-150 bp, followed by end repair and addition of a 3’ A base. Cytosine-methylated adapters provided by Illumina (Illumina, San Diego, CA) were ligated to the sonicated DNA at 16˚C for 16 hours with T4 DNA ligase (New England Biolabs). Adapter-ligated DNA was isolated by two rounds of purification with AMPure XP beads (Beckman Coulter Genomics, Danvers, MA). Adapter-ligated DNA (≤450 ng) was subjected to sodium bisulfite conversion using the MethylCode kit (Life Technologies, Carlsbad, CA) as per manufacturer’s instructions. The bisulfite-converted, adapter-ligated DNA molecules were enriched by 4 cycles of PCR with the following reaction composition: 2.5 U of uracil-insensitive PfuTurboCx Hotstart DNA polymerase (Stratagene), 5 µl 10X PfuTurbo reaction buffer, 31 µM dNTPs, 1 µl Primer 1, 1 µl Primer 2 (50 µl final). The thermocycling parameters were: 95˚C 2 min, 98˚C 30 sec, then 4-8 cycles of 98˚C 15 sec, 60˚C 30 sec and 72˚C 4 min, ending with one 72˚C 10 min step. The reaction products were purified using AMPure XP beads (two rounds). Up to three separate PCR reactions were performed on subsets of the adapter-ligated, bisulfite-converted DNA, yielding up to two independent libraries from the same biological sample.
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| Library strategy |
Bisulfite-Seq |
| Library source |
genomic |
| Library selection |
RANDOM |
| Instrument model |
Illumina Genome Analyzer II |
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| Description |
Design description: Whole genome shotgun bisulfite sequencing of the H1 cell line treated with BMP4 for 5 days
Library name: methylC-seq_h1+bmp4_r2a
EDACC Genboree Experiment Page:
http://genboree.org/java-bin/project.jsp?projectName=XML%20Submissions%2FUCSD%2FEXPERIMENT%2FEDACC.3382
EDACC Genboree Sample Page:
http://genboree.org/java-bin/project.jsp?projectName=XML%20Submissions%2FUCSD%2FSAMPLE%2FEDACC.3378
****************
For data usage terms and conditions, please refer to:
http://www.drugabuse.gov/funding/funding-opportunities/nih-common-fund/epigenomics-data-access-policies
****************
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| Data processing |
**********************************************************************
ANALYSIS FILE NAME: GSM602254_UCSD.H1-BMP4.Bisulfite-Seq.methylC-seq_h1+bmp4_r2a.wig
ANALYSIS CENTER: EDACC
ANALYSIS ALIAS: methylC-seq_h1+bmp4_r2a.hg19.level.2
ANALYSIS TITLE: Methylation Proportion Graphs of H1-BMP4 Cell Line Bisulfite-Seq Data
ANALYSIS DESCRIPTION: Illumina Bisulfite-Seq read mappings from the H1-BMP4 Cell Line, Library methylC-seq_h1+bmp4_r2a were processed into graphs of methylation proportions. Methylation proportions were calculated as (methylated calls / (methylated calls + unmethylated calls)) for all CpGs covered by at least 4 reads. Reads from the + and - strands were combined for methylation proportion calculations.
ANALYSIS TYPE: ABUNDANCE_MEASUREMENT
EDACC Genboree Analysis Page: http://genboree.org/java-bin/project.jsp?projectName=XML%20Submissions%2FEDACC%2FANALYSIS%2FEDACC.5122
DATA_ANALYSIS_LEVEL: 2
EXPERIMENT_TYPE: Bisulfite-Seq
GENOME_ASSEMBLY: NCBI Build GRCh37/UCSC Build hg19
SOFTWARE: In house programs and scripts
SOFTWARE_VERSION: NA
READ_EXTENSION: 0bp
TREATMENT_OF_IDENTICAL_ALIGNMENTS_OF_MULTIPLE_READS: If multiple reads map to the same start position on the + strand or stop position on the - strand, only a single read is retained.
GENOMIC_WINDOW: 2bp containing CpGs
TREATMENT_OF_REGIONS_PRONE_TO_MULTIPLE_ALIGNMENTS: None
RELEASE_NUMBER: Human Epigenome Atlas 2
BROWSER_TRACK_NAME: H1BMP4 BS 2a
BROWSER_TRACK_DESCRIPTION: UCSD H1-BMP4 Cell Line Bisulfite-Seq Library methylC-seq_h1+bmp4_r2a EA Release 2
QUALITY SCORES:
NUMBER_OF_Bisulfite-Seq_EXPERIMENTS_SCORED: 17
BISULFITE_CONVERSION_PERCENTAGE_BASED_ON_MAPPINGS: 98.47
BISULFITE_CONVERSION_PERCENTAGE_BASED_ON_MAPPINGS_PERCENTILE: 65
**********************************************************************
ANALYSIS FILE NAME: GSM602254_UCSD.H1-BMP4.Bisulfite-Seq.combined.wig
ANALYSIS CENTER: EDACC
ANALYSIS ALIAS: methylC-seq_h1+bmp4_r1a-methylC-seq_h1+bmp4_r1b-methylC-seq_h1+bmp4_r1c-methylC-seq_h1+bmp4_r2a-methylC-seq_h1+bmp4_r2b-methylC-seq_h1+bmp4_r2c.hg19.level.2
ANALYSIS TITLE: Methylation Proportion Graphs of H1-BMP4 Cell Line Bisulfite-Seq Data
ANALYSIS DESCRIPTION: Illumina Bisulfite-Seq read mappings from the H1-BMP4 Cell Line combined libraries were processed into graphs of methylation proportions. Methylation proportions were calculated as (methylated calls / (methylated calls + unmethylated calls)) for all CpGs covered by at least 4 reads. Reads from the + and - strands were combined for methylation proportion calculations.
ANALYSIS TYPE: ABUNDANCE_MEASUREMENT
EDACC Genboree Analysis Page: http://genboree.org/java-bin/project.jsp?projectName=XML%20Submissions%2FEDACC%2FANALYSIS%2FEDACC.5129
DATA_ANALYSIS_LEVEL: 2
EXPERIMENT_TYPE: Bisulfite-Seq
GENOME_ASSEMBLY: NCBI Build GRCh37/UCSC Build hg19
SOFTWARE: In house programs and scripts
SOFTWARE_VERSION: NA
READ_EXTENSION: 0bp
TREATMENT_OF_IDENTICAL_ALIGNMENTS_OF_MULTIPLE_READS: If multiple reads map to the same start position on the + strand or stop position on the - strand, only a single read is retained.
GENOMIC_WINDOW: 2bp containing CpGs
TREATMENT_OF_REGIONS_PRONE_TO_MULTIPLE_ALIGNMENTS: None
RELEASE_NUMBER: Human Epigenome Atlas 2
BROWSER_TRACK_NAME: H1BMP4 BS Combined
BROWSER_TRACK_DESCRIPTION: UCSD H1-BMP4 Cell Line Bisulfite-Seq Combined Libraries EA Release 2
BISULFITE_CONVERSION_PERCENTAGE_BASED_ON_MAPPINGS: 98.32
SPECIAL NOTE: generated from combined data in GSM602251, GSM602252, GSM602253, GSM602254, GSM602255, GSM602256 (thus .wig files on these records are identical)
**********************************************************************
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| Submission date |
Sep 29, 2010 |
| Last update date |
May 15, 2019 |
| Contact name |
UCSD AND SALK |
| Organization name |
University of California, San Diego
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| Street address |
Health Sciences Drive
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| City |
La Jolla |
| State/province |
CA |
| ZIP/Postal code |
92092 |
| Country |
USA |
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| Platform ID |
GPL9115 |
| Series (1) |
| GSE16256 |
UCSD Human Reference Epigenome Mapping Project |
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| Relations |
| SRA |
SRX027688 |
| BioSample |
SAMN00114947 |
| Named Annotation |
GSM602254_UCSD.H1-BMP4.Bisulfite-Seq.methylC-seq_h1+bmp4_r2a.wig.gz |
| Named Annotation |
GSM602254_UCSD.H1-BMP4.Bisulfite-Seq.combined.wig.gz |
| Supplementary file |
Size |
Download |
File type/resource |
| GSM602254_UCSD.H1-BMP4.Bisulfite-Seq.combined.wig.gz |
229.0 Mb |
(ftp)(http) |
WIG |
| GSM602254_UCSD.H1-BMP4.Bisulfite-Seq.methylC-seq_h1+bmp4_r2a.wig.gz |
153.3 Mb |
(ftp)(http) |
WIG |
SRA Run Selector |
| Processed data provided as supplementary file |
| Raw data are available in SRA |
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