GSM1847652: MIRA-6543; Homo sapiens; OTHER
Genomic DNAs from human melanocytes were prepared using Quick-gDNA MiniPrep kit (Zymo Research, Cat# D3007) followed the manufacture's method. For Methylated-CpG island recovery assay (MIRA), two micrograms of genomic DNA was sonicated to produce DNA fragments with a length distribution of 100 to 300 bp using a Covaris S220 sonicator. The sheared DNA was Illumina linker-ligated, and methylated DNA fragments were enriched using the methylated-CpG island recovery assay (MIRA) technique using recombinant MBD3L1 and MBD2b proteins. To prepare a sequencing library, the MIRA-enriched DNA fragments and input DNA were PCR-enriched with PE 1.0 primer and index primer (Illumina). Next, Ampure XP beads (Beckman Coulter; Brea, CA) was added to the PCR solution to remove DNA fragments smaller than 200 bp, and double bead selection (0.65x) was used to remove fragments larger than 500 bp according to the manufacturer’s instructions. The size-selected sequencing library was sequenced on an Illumina HiSeq instrument. MIRA-Seq
GEO
GSM1847652