The length of telomeric G-rich strand 3'-overhang measured by oligonucleotide ligation assay.

G. Cimino-Reale, E. Pascale, E. Battiloro, G. Starace, R. Verna, E. D'Ambrosio

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Abstract

A typical G-rich telomeric DNA strand, which runs 5'-->3' toward the chromosome ends, protrudes by several nucleotides in lower eukaryotes. In human chromosomes long G-rich 3'-overhangs have been found. Apart from the standard G-rich tail, several non-canonical terminal structures have been proposed. However, the mechanism of long-tail formation, the presence and the role of these structures in telomere maintenance or shortening are not completely understood. In a search for a simple method to accurately measure the 3'-overhang we have established a protocol based on the ligation of telomeric oligonucleotide hybridized to non-denatured DNA under stringent conditions (oligonucleotide ligation assay with telomeric repeat oligonucleotide). This method enabled us to detect a large proportion of G-rich single-stranded telomeric DNA that was as short as 24 nt. Nevertheless, we showed G-tails longer than 400 nt. In all tested cells the lengths ranging from 108 to 270 nt represented only 37% of the whole molecule population, while 56-62% were

Original languageEnglish
JournalNucleic Acids Research
Volume29
Issue number7
Publication statusPublished - Apr 1 2001

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ASJC Scopus subject areas

  • Genetics

Cite this

Cimino-Reale, G., Pascale, E., Battiloro, E., Starace, G., Verna, R., & D'Ambrosio, E. (2001). The length of telomeric G-rich strand 3'-overhang measured by oligonucleotide ligation assay. Nucleic Acids Research, 29(7).