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Nucleic Acids Res. 2014 Feb;42(3):e21. doi: 10.1093/nar/gkt1315. Epub 2013 Dec 23.

TeloTool: a new tool for telomere length measurement from terminal restriction fragment analysis with improved probe intensity correction.

Author information

1
Max F. Perutz Laboratories, Medical University of Vienna, Vienna 1030, Austria, Gregor Mendel Institute, Vienna 1030, Austria, Institute for Applied Physics, Johannes Kepler University Linz, Linz 4040, Austria and Upper Austria University of Applied Sciences, Campus Linz, Linz 4020, Austria.

Abstract

Telomeres comprise the protective caps of natural chromosome ends and function in the suppression of DNA damage signaling and cellular senescence. Therefore, techniques used to determine telomere length are important in a number of studies, ranging from those investigating telomeric structure to effects on human disease. Terminal restriction fragment (TRF) analysis has for a long time shown to be one of the most accurate methods for quantification of absolute telomere length and range from a number of species. As this technique centers on standard Southern blotting, telomeric DNA is observed on resulting autoradiograms as a heterogeneous smear. Methods to accurately determine telomere length from telomeric smears have proven problematic, and no reliable technique has been suggested to obtain mean telomere length values. Here, we present TeloTool, a new program allowing thorough statistical analysis of TRF data. Using this new method, a number of methodical biases are removed from previously stated techniques, including assumptions based on probe intensity corrections. This program provides a standardized mean for quick and reliable extraction of quantitative data from TRF autoradiograms; its wide application will allow accurate comparison between datasets generated in different laboratories.

PMID:
24366880
PMCID:
PMC3919618
DOI:
10.1093/nar/gkt1315
[Indexed for MEDLINE]
Free PMC Article

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