GSM2087091: two-weeks seedling plants_308D_MeDIPseq; Oryza sativa; MeDIP-Seq
For each sample, about 20 individual plants were pooled and total genomic DNA was extracted using CTAB method. DNA samples were treated with RNase A and DNA was purified from the supernatant using chloroform.For each sample, about 20 individual plants were pooled and total genomic DNA was extracted using CTAB method. DNA samples were treated with RNase A and DNA was purified from the supernatant using chloroform.  About 5 microgram of genomic DNA was sheared by sonication to fragments of 100-500 bp. Sonicated DNA was end-repaired, A-tailed, and ligated to single-end adapters following the standard Illumina protocol. After agarose size-selection to remove unligated adapters, 2–5 μg of adaptor-ligated DNA was used for each immunoprecipitation using a mouse monoclonal anti-methylcytidine antibody, following performed by Magnetic Methylated DNA Immunoprecipitation kit (Diageno,Catalog No:  mc-magme-048). MeDIP DNA was purified using Qiagen MinElute columns and eluted in 16 μl EB (Qiagen). Fifteen cycles of PCR were performed on 5 μl of the immunoprecipitated DNA using the single-end Illumina PCR primers. Libraries were quality controlled by spectrophotometry and Agilent DNA Bioanalyzer analysis. Libraries were sequenced on the Hiseq2000 following the manufacturer's protocols. libraries were prepared for sequencing using standard Illumina protocols.
GEO Accession
GSM2087091