GSM3666845: Liver Frozen K27Ac; Mus musculus; ChIP-Seq
Lysates were clarified from Mnase/sonicated nuclei and histone-DNA complexes were isolated with antibody. Libraries were prepared according to ThruPLEX® DNA-seq Kit V2 (Rubicon Genomics, Ann Arbor, MI) according to the manufacturer's instructions. . Briefly,In the first step, the DNA is repaired and yields molecules with blunt ends. In the next step, stem-loop adaptors with blocked 5' ends are ligated with high efficiency to the 5' end of the genomic DNA, leaving a nick at the 3' end. The adaptors cannot ligate to each other and do not have single-strand tails, both of which contribute to non-specific background found with many other NGS preparations. In the final step, the 3' ends of the genomic DNA are extended to complete library synthesis and Illumina-compatible indexes are added through a high-fidelity amplification. After adapter ligation DNA was PCR amplified with Illumina primers for 12 cycles and library fragments of ~300 bp (insert plus adaptor and PCR primer sequences) were purified by AMpure reagents. The purified DNA was captured on an Illumina flow cell for cluster generation. Libraries were sequenced on the Illumina HiSeq 4000 following the manufacturer's protocols.
GEO
GSM3666845