DAP-seq of ERF.F12_2 (CRX1925195; CRA029190)
(1) Genomic DNA was extracted from tissue materials, fragmented to approximately 200bp, sequenced with a linker and amplified by PCR, and finally a linker amplified DNA library (ampDAP library) was prepared;  (2) Clone the target transcription factor into the expression vector, synthesize the fusion protein (TF-Halo) with an affinity tag (such as Halo or GST) through the in vitro expression system, and then immobilized through affinity magnetic beads;  (3) Affinity purification and library construction: Immobilized transcription factors are co-incubated with ampDAP libraries to specifically bind to their target DNA sequences (TFBS). After elution of unbound DNA, the bound DNA fragments are subjected to PCR amplification to construct sequencing libraries for the identification of transcription factor binding sites across the entire genome.
NGDC
CRX1925195
Southwest University of Science and Technology
DRX812096