DNA from human keratinocyte cells was extracted using the GenElute Mammalian Genomic DNA Miniprep Kit (Sigma-Aldrich) according to manufacturer’s instructions. DNA was nicked using the Nt.BspQI enzyme (10,000 U/mL; NEB). 10 μg of DNA were mixed with 11.6 μL Nt.BspQI enzyme, 10 μL NEBuffer 3.1 (NEB), and ultrapure water to a final volume of 70 μL. The reaction mixture was incubated for 2 h at 50°C. After the nicking reaction, DNA samples were labeled with 10 μL of a cocktail of repair enzymes (PreCR Repair Mix; NEB), fragmented, and assayed according to RADD-seq procedure as described above, with biotinylated nucleotides.
Nt bspqi enzyme
Manufactured by New England Biolabs
Nt.BspQI is a nicking endonuclease enzyme that recognizes and cleaves a specific DNA sequence, creating a single-strand break. The enzyme functions independently of DNA methylation status.
Automatically generated - may contain errors
1 protocol using nt bspqi enzyme
1
DNA Nicking and RADD-seq Profiling
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