Methylated DNA Immunoprecipitation Sequencing of AC7729/TZSRW . Not stressed .
Purified DNA was sent to BGI for MeDIP-seq analysis followed by the protocol in the previous study (Taiwo et al. 2012). Briefly, DNA sonicated into fragments ranging from 100–500 bp were retracted with gel extraction kit (Qiagen) and ligated with an Illumina sequencing primer adapter after end repairing, phosphorylating and A-tailing with Paired-End DNA Sample Prep kit (Illumina, San Diego, CA, USA). Then the fragments were used for MeDIP enrichment using Magnetic Methylated DNA Immunoprecipitation kit (Diagenod, Liège, Belgium) following the manufacturer’s recommendation and the qualifying DNA was used for PCR amplification. Then bands between 220 and 320 bp were excised from the gel and purified with QIAquick Gel Extraction Kit (Qiagen, Valencia, CA, USA). The eight DNA libraries were sequenced on the Illumina Hiseq 2000 (Illumina, San Diego, CA, USA) to generate paired-end 50-bp reads by the Beijing Genomics Institute (BGI, Shenzhen, Guangdong, China).