A practical guide to studying genome function using single-molecule genomics.

Single-molecule genomics (SMG) has transformed our ability to study the mechanisms that regulate the genome by enabling profiling of the activity of regulatory factors on individual DNA molecules genome-wide. SMG is able to quantify molecular heterogeneity and the co-occurrence of regulatory events, including epigenetic modifications, transcription factor binding and chromatin organization on single DNA molecules. SMG reveals dynamics of chromatin interactions that cannot be measured by conventional genomics assays. Therefore, SMG offers a unique platform to study how regulatory events combine to control genome activity. In this Expert Recommendation article, we provide a practical guide for adopting SMG and outline best practices.