GSM6601356: NCP-H3K27me3-NRCAM-OCT4-Round4-Rep1; synthetic construct; OTHER
Sequencing libraries were prepared by adding to the selected libraries (1) 5' end Unique Molecular Identifies (UMIs), (2) llumina adaptors (IAs), (2) TRUseq dual barcodes (BCs) and (2) Illumina sequencing promoters P5 and P7 (P5/P7) by PCR amplification with custom primers.  10 ng of DNA was used as starting material samples. Libraries were with KAPA HiFi HotStart ReadyMix PCR Kit amplified by using 2 consecutive 6-cycle PCR runs (first cycle for UMIs and IAs, second cycle for BCs and P5/P7)  on the thermocycler. Post amplification libraries were size selected at ~3000bp in length by Azenta using AMPure XP beads. Libraries were validated by using both the Tapestation D1000 and  Qubit. Library were selected 4 times by: assembly on Nucleosome core particle, incubation with GST-tagged TFs, immunoprecipiation by GST beads and PCR amplification.