Quantitative telomeric overhang determination using a double-strand specific nuclease

Telomeres terminate in 3′ overhangs that function in end protection and the formation of t-loops. Determining the steps and factors involved in overhang processing is compromised by the inability to easily and accurately determine overhang size in the presence of many kilobases of double-stranded telomeric DNA. We here describe the use of a double-strand specific nuclease (DSN) that entirely digests double-stranded DNA including telomeres, leaving the overhangs intact so that they can be measured.