Zhao Yong, Shay Jerry W, Wright Woodring E
Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, TX, USA.
Methods Mol Biol. 2011;735:47-54. doi: 10.1007/978-1-61779-092-8_5.
Telomeres terminate in 3' single-stranded G-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 genomic DNA including telomeres, leaving the single-stranded overhangs intact. The telomere single-strand G-rich overhang length can then be determined by Southern blot-based assays.
端粒以3'单链G突出端结束，这些突出端在端粒末端保护和端粒酶作用中发挥功能。由于存在数千碱基对的双链端粒DNA，准确测量突出端长度具有挑战性。在此，我们描述了一种简单的方法，该方法利用双链特异性核酸酶（DSN）消化包括端粒在内的所有基因组DNA，使单链突出端保持完整。然后可以通过基于Southern印迹的分析来确定端粒单链富含G的突出端长度。
