使用双链特异性核酸酶进行端粒悬垂定量测定。

Zhao Yong, Hoshiyama Hirotoshi, Shay Jerry W, Wright Woodring E

Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, Texas, USA.

Nucleic Acids Res. 2008 Feb;36(3):e14. doi: 10.1093/nar/gkm1063. Epub 2007 Dec 10.

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.

端粒以3'端突出端结束，其在末端保护和t环形成中发挥作用。由于在存在许多千碱基的双链端粒DNA的情况下难以轻松准确地确定突出端大小，因此确定突出端加工所涉及的步骤和因素受到了阻碍。我们在此描述了一种双链特异性核酸酶（DSN）的用途，该酶可完全消化包括端粒在内的双链DNA，使突出端保持完整以便进行测量。