GSM5331362: Cop1_sgRNA#2_Cebpd_ChIP_seq_R2_treat_pileup.bw; Mus musculus; ChIP-Seq
Lysates were clarified from sonicated nuclei and Cebpd-DNA complexes were pulled down with antibody. Libraries were prepared according toTAKARA's instructions accompanying Smarter Thruplex DNAseq Kit. Briefly, in the first step, Template Preparation, 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 can not 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. Any remaining free adaptors are destroyed. Hands-on time and the risk of contamination are minimized by using a single tube and eliminating intermediate purifications.