Analysis of Telomere-Homologous DNA with Different Conformations Using 2D Agarose Electrophoresis and In-Gel Hybridization.
In mammalian cells, in addition to double-stranded telomeric DNA at chromosome ends, extra telomere-homologous DNA is present that adopts different conformations, including single-stranded G- or C-rich DNA, extrachromosomal circular DNA (T-circle), and telomeric complex (T-complex) with an unidentified structure. The formation of such telomere-homologous DNA is closely related to telomeric DNA metabolism and chromosome end protection by telomeres. Conventional agarose gel electrophoresis is unable to separate DNA based on conformation. Here, we introduce the method of two-dimensional (2D) agarose electrophoresis in combination with in-gel native/denatured hybridization to determine different conformations formed by telomere-homologous DNA.
Key Laboratory of Gene Engineering of the Ministry of Education, Higher Education Mega Center, School of Life Sciences, Sun Yat-sen University, 132 East Waihuan Rd., Guangzhou, 510006, P. R. China.
Zepeng
Z
Zhang
Key Laboratory of Gene Engineering of the Ministry of Education, Higher Education Mega Center, School of Life Sciences, Sun Yat-sen University, 132 East Waihuan Rd., Guangzhou, 510006, P. R. China.
Qian
Q
Hu
Key Laboratory of Gene Engineering of the Ministry of Education, Higher Education Mega Center, School of Life Sciences, Sun Yat-sen University, 132 East Waihuan Rd., Guangzhou, 510006, P. R. China. zhaoy82@mail.sysu.edu.cn.
Yong
Y
Zhao
D000818:Animals; D002875:Chromosomes / Q000235:genetics; D004270:DNA, Circular / Q000235:genetics*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D009693:Nucleic Acid Hybridization / Q000235:genetics*; D019098:Telomerase / Q000235:genetics; D016615:Telomere / Q000235:genetics*
D004270:DNA, Circular; D019098:Telomerase
2D agarose electrophoresis; DNA conformations; In-gel hybridization; Telomere