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Sample GSM2319712 Query DataSets for GSM2319712
Status Public on Sep 18, 2016
Title wild-type postmature-green-stage seed
Sample type SRA
 
Source name postmature-green-stage seeds from Arabidopsis wild-type
Organism Arabidopsis thaliana
Characteristics genotype: wild type
tissue: seed
cultivar: Col
Growth protocol Arabidopsis thaliana (L.) Heynh, ecotype Columbia (Col) wild type and ddcc mutant plants were grown under standard greenhouse conditions (Belmonte et al., PNAS 2013). Leaves were collected for genomic DNA isolation from 3 week plant.
Extracted molecule genomic DNA
Extraction protocol Collected seeds and leaves were quickly frozen in liquid nitrogen and ground to a fine powder using a mortar and pestle. Genomic DNA was isolated from the powder using the DNEASY Plant Mini kit (Qiagen, Valencia, CA) according to manufacturer’s instructions.
Approximately 500 nanogram of genomic DNA isolated from seeds and leaves was subjected to library preparation following the methods of Hsieh et al. (Hsieh T-F et al. 2009. Science 324:1451-1454) with modifications. We spiked-in ~ three nanogram of unmethylated lambda DNA (Promega) to serve as a control for complete bisulfite conversion. Adapter-ligated genomic DNA was subjected to two rounds of bisulfite (BS) treatment using the EpiTect kit (Qiagen, Valencia, CA). BS-treated DNA was purified using AMpure XP beads (Beckman) and PCR-amplified for 10 cycles using ExTaq (EpiCentre) DNA polymerase. PCR-amplified DNA fragments were size selected using the AMpure XP beads (Beckman). Phi-X174 DNA was spiked in to the library by the sequencing facility before cluster formation and sequencing.
 
Library strategy Bisulfite-Seq
Library source genomic
Library selection RANDOM
Instrument model Illumina HiSeq 2000
 
Description Postmature-green-stage seeds from Arabidopsis Col wild-type were collected
Data processing Basecalls performed using RTA version 1.12.4.2
Original data file from the Illumina sequencing pipeline. Each lane of sequencing is attached as an individual compressed file. The sequence data are in the QSEQ format
We aligned the raw reads to a pre-processed reference genome using BS Seeker [Chen et al. BMC Bioinformatics (2010)] allowing for two mismatches. The pre-processed reference genome consisted of sequences from the Arabidopsis thaliana genome (TAIR 10) obtained from the TAIR website (http://www.arabidopsis.org/) and Phi-X174 reference genome (GenBank: J02482).
Reads containing three consecutive methylation in the non-CG sites were removed, possibly representing non-converted cytosines (Cokus et al. Nature 2008). Clonal reads possibly arising during the PCR amplification step were collapsed into one read.
Methylation level of sampled cytosine was calculated as (methylated calls / (methylated calls + unmethylated calls))
Genome_build: TAIR 10
Supplementary_files_format_and_content: tab-delimited text file including methylation level of each sampled cytosine
 
Submission date Sep 17, 2016
Last update date Sep 18, 2016
Contact name Bob Goldberg
E-mail bobglab@mcdb.ucla.edu
Phone 310-825-3270
Organization name University of California, Los Angeles
Department Molecular, Cell and Developmental Biology
Street address 610 Charles E Young Drive East
City Los Angeles
State/province CA
ZIP/Postal code 90095
Country USA
 
Platform ID GPL13222
Series (1)
GSE68131 Methylation Profile of postmature-green-stage seed and dry seed from Arabidopsis ddcc mutant
Relations
BioSample SAMN05785628
SRA SRX2173461

Supplementary file Size Download File type/resource
GSM2319712_At.Col0.WT.wm.pmg.seed.bsseq.chr1-5.perC.txt.gz 168.6 Mb (ftp)(http) TXT
Raw data provided as supplementary file
Processed data provided as supplementary file

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