2019/04/18 by Hatice S Kaya-Okur, Steven Henikoff · 1 citation
Biochemistry, Genetics and Molecular Biology · #Advanced biosensing and bioanalysis techniques #Genomics and Chromatin Dynamics #Cancer Genomics and Diagnostics
paper · pdf · doi:10.17504/protocols.io.z6hf9b6
openalex publication_date 2019/04/18 · openalex created_date 2022/05/12 · openalex updated_date 2026/07/14
Here we describe a single-tube bench top protocol for Cleavage Under Targets and Tagmentation (CUT&Tag), an enzyme-tethering strategy that provides efficient high-resolution sequencing libraries for profiling diverse chromatin components. In CUT&Tag, a chromatin protein is bound in situ by a specific antibody, which then tethers a proteinA-Tn5 transposase fusion protein. Activation of the transposase efficiently generates fragment libraries with high resolution and exceptionally low background. All steps from live cells to sequencing-ready libraries can be performed in a single tube on the benchtop or a microwell in a high-throughput pipeline, and the entire procedure can be performed in one day. We have demonstrated the utility of CUT&Tag by profiling histone modifications, RNA Polymerase II and transcription factors on low cell numbers and single cells.