RAP2-7 DAP-seq (Lupinus angustifolius L.)
The immobilized Halo-tagged allele-specific RAP2-7 protein was incubated with the adapter-ligated DNA library. After incubation, the beads were washed, and the bound DNA was recovered, PCR-amplified, purified, and quantified. Negative controls were processed in parallel using adapter-ligated DNA libraries incubated with Magne HaloTag beads in the absence of recombinant protein. For more details please see accompanying manuscript.  Plants were cultivated in a growth chamber (16 h photoperiod, 22/18 °C day/night, 55–60% relative humidity, until they reached whole plant flowering stage. High-molecular-weight genomic DNA for DAP-seq was extracted from frozen young leaves (3 g) using a phenol:chloroform:isoamyl alcohol protocol, and quantified fluorometrically using the Qubit dsDNA Broad Range Assay (Thermo Fisher Scientific). Genomic DNA libraries were prepared according to Bartlett et al. (2017; doi.org/10.1038/nprot.2017.055), with minor modifications. For more details please see accompanying manuscript.