The protein-oligo conjugate bands were excised from the SDS-PAGE gel, and the protein-oligo complex were recovered from gel slab. Two rounds of PCR amplification were carried out for next-generation sequencing library preparation. 10% of the materials recovered from the gel was used for PCR amplification. First, the DNA part of the protein-DNA conjugate was amplified using its PCR handle. Thensequencing adapters were appended using NEB Next Multiplex Oligos for Illumina (NEB) in second-round PCR. After each round of PCR, PCR amplicons were separated on 3% agarose gel and purified using gel extraction kit (Thermo Scientific, K210012) without heating.
GEO Accession
GSM4647879