Telomere G-Rich Overhang Length Measurement: DSN Method.
Telomeres terminate in 3' single-stranded G-rich overhangs that function in telomere end protection and telomerase action. An accurate measurement of overhang length is challenging due to the presence of many kilobases of double-stranded telomere DNA. Here, a simple method is described that utilizes duplex specific nuclease (DSN) to digest all double-stranded DNA including telomeres to fragments of <10 bp, leaving the single-stranded overhangs intact. Their length can then be determined by Southern blot using super-sensitive telomere C-rich probes.
Key Laboratory of Gene Engineering of the Ministry of Education, School of Life Sciences, Sun Yat-sen University, Guangzhou, 510006, People's Republic of China.
Yong
Y
Zhao
Department of Cell Biology, University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd., Dallas, TX, 75390, USA.
Jerry W
JW
Shay
Department of Cell Biology, University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd., Dallas, TX, 75390, USA. Woodring.Wright@UTSouthwestern.edu.
Woodring E
WE
Wright
D000818:Animals; D015139:Blotting, Southern / Q000379:methods; D002478:Cells, Cultured; D004247:DNA / Q000235:genetics; D004261:DNA Replication / Q000235:genetics*; D004277:DNA, Single-Stranded / Q000235:genetics; D003851:Deoxyribonucleases / Q000235:genetics*; D008322:Mammals; D019098:Telomerase / Q000235:genetics; D016615:Telomere / Q000235:genetics*
D004277:DNA, Single-Stranded; D004247:DNA; D019098:Telomerase; D003851:Deoxyribonucleases
Duplex specific nuclease (DSN); G-rich overhang; Southern blot; Telomere; Telomere C-rich probe