|
Status |
Public on May 11, 2016 |
Title |
F0H MeDIP |
Sample type |
SRA |
|
|
Source name |
F0H
|
Organism |
Oryzias melastigma |
Characteristics |
group: Hypoxic group in F0 generation cell type: sperm
|
Extracted molecule |
genomic DNA |
Extraction protocol |
For each group, sperm from 10 male fish were collected by gently pressing the harvested testis and pooled. Sperm genomic DNA was extracted using a DNA extraction kit (MO BIO Ultraclean® Tissue & Cells DNA Isolation Kit, USA). Extracted DNA was prepared by the Paired-End DNA Sample Prep Kit (Illumina) according to the manufacturer’s instructions. Briefly, DNA was sonicated to fragments of 100-500 bp before the addition of dAs and adaptor ligation. Methylated DNA was then pulled down by an antibody specifically recognizing 5-methylcytosine. The pull-down fraction containing the antibody-5-methylcytosine complex was subjected to column purification (Zymo Research). The purified DNA was next enriched by an adaptor-mediated PCR process. Fragments of 220-232 bp were gel purified with a Gel Extraction Kit (Qiagen).
|
|
|
Library strategy |
MeDIP-Seq |
Library source |
genomic |
Library selection |
5-methylcytidine antibody |
Instrument model |
Illumina HiSeq 2000 |
|
|
Data processing |
The genome sequence of freshwater medaka was used as the reference genome for bowtie alignment. DiffReps software was employed to identify the differentially methylated regions with a FDR cut-off of 0.01. Genome_build: Oryzias_latipes release 73 downloaded from ensembl Supplementary_files_format_and_content: .wig files report peaks
|
|
|
Submission date |
May 11, 2016 |
Last update date |
May 15, 2019 |
Contact name |
Eric Greer |
E-mail(s) |
Eric.Greer@childrens.harvard.edu
|
Organization name |
Boston Children's Hospital/Harvard Medical School
|
Department |
Newborn Medicine/Department of Pediatrics
|
Lab |
Greer Lab
|
Street address |
320 Longwood Avenue
|
City |
Boston |
State/province |
MA |
ZIP/Postal code |
02115 |
Country |
USA |
|
|
Platform ID |
GPL21858 |
Series (1) |
GSE81312 |
MeDIP on the marine medaka sperm (Oryzias melastigma) |
|
Relations |
BioSample |
SAMN04987094 |
SRA |
SRX1756769 |