Name: GSM8023945
Instrument: NextSeq 2000
Strategy: MeDIP-Seq
Source: GENOMIC
Selection: 5-methylcytidine antibody
Layout: PAIRED
Construction protocol: DNA was extracted from 6 biological replicates using DNA Easy Blood and Tissue Kit (Qiagen, 69506) according to instructions provided by manufacturer. DNA concentration was measured by fluorescent method using fluorescent reader (Promega). 10 µgs of DNA was used for MeDIP using methylated DNA Enrichment Kit (NEB, #E2600S). DNA was diluted and sonicated in Qsonica sonicator using following conditions: 20 s pulse on, 20 sec pulses off, total sonication time is 8 min. After that, sonicated DNA was incubated with MBD2-Fc -protein A Magnetic beads. The beads were washed 4 times with 1XBind/Wash Buffer. The enriched methylated DNA was eluted from beads with miliQ water and and the concentration of methylated DNA was again determined by fluorescence. 6 ng of methylated DNA or input were taken for library preparation. Sequencing libraries were prepared using NEBNext Ultra DNA Library Prep Kit for Illumina (E7645S; NEB). We performed 15 cycles for the library amplification.