GSM4483713: CNTR 2 [MeDIP-seq]; Mus musculus; MeDIP-Seq
Genomic DNA was extracted with Diagenode kit (MagMeDIP kit; Cat nr: C15200081), sonicated with the Bioruptor, and precipitated with 5meC-Ab (Diagenode; Cat nr: C15200081; Lot nr: RD004) or with control IgG (Diagenode; Cat nr: C15400001; Lot nr: MIG002S). MeDIP-seq libraries were generated from 1 to 4 ng of single-stranded or double-stranded IP-DNA samples using the DNA SMART ChIP-Seq (634866, Takara Bio USA, Inc., Mountain View, CA, USA) according to manufacturer's instructions. Briefly, the differents steps of the library prep were: 1) denaturation of the DNA, 2) T-tailing of the 3' end of the single-stranded DNA by the terminal transferase 3), annealing of DNA SMART poly(dA) primer to the 3' T-tail, 4) primer extension by the reverse transcriptase (RT), 5) template switching and extension by RT using SMART oligo, 6) addition of Illumina indexed adapters through a PCR amplification (12 cycles) using primers compatible with regions on the DNA SMART Poly(dA) primer and the SMART oligo, 7) purification and size-selection of amplified library using Agencourt AMPure XP beads (Beckman Coulter) to remove unincorporated primers and other reagents.
GEO Accession
GSM4483713