LABEsC1C1
1g genomic DNA was digested using MspI enzyme for 16 hours at 37. After digestion, libraries were constructed as the Illumina Pair-End protocol with some modifications. Briefly, purified digested DNA was subsequently treated with a mix of T4 DNA polymerase, Klenow Fragment and T4 polynucleotide kinase to repair, blunt and phosphorylate ends. After that, sequencing control DNA supplied by the TrueMethyl Seq Kit (CEGX) were mixed with the blunt DNA. The mixture DNA fragments were subsequently 3 adenylated using Klenow Fragment (3-5 exo-) and following with ligation to adaptors synthesized with 5-methylcytosine instead of cytosine using T4 DNA Ligase. the DNA was purified using QIAquick PCR purification kit (Qiagen) after reaction of each step.  Before oxidation reaction, all products had been purified using Magnetic Beads Binding Solution 1 supplied by the TrueMethyl Seq Kit (CEGX) according to the manufactures instructions. After purification, the oxidation reaction was conducted in a reaction volume of 25 l with 1 l of the Oxidant Solution (CEGX) for each oxRRBS library, and RRBS library sample added in 1ul of ultrapure water instead of Oxidant as control. Both of the two libraries were subjected to 40 C for 30 min treatment in a thermo cycler with the lid heated at 57C. After that, centrifuged the reaction mixture at 14,000 X g for 10 min and then transferred the supernatant into a new 0.2 ml PCR tube for the further bisulfite treatment, respectively. Bisulfite conversion treatment was performed using a TrueMethyl Seq Kit  (CEGX) according to the manufactures instructions. The final oxRRBS and RRBS libraries were generated by PCR amplification using adapter compatible barcode primers, quantified by an Agilent 2100 Bioanalyzer (Agilent Technologies) and real-time PCR assay and then sequenced by Illumina sequencing platform.