| VALUCHOVÁ, Soňa, Elisa DERBOVEN a Karel ŘÍHA. Analysis of Telomeric G -overhangs by in- Gel Hybridization. Bio-protocol. Sunnyvale: Bio-protocol LLC, 2016, roč. 6, č. 7, s. nestránkováno, 9 s. ISSN 2331-8325. Dostupné z: https://dx.doi.org/10.21769/BioProtoc.1775. |
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@article{1376223,
author = {Valuchová, Soňa and Derboven, Elisa and Říha, Karel},
article_location = {Sunnyvale},
article_number = {7},
doi = {http://dx.doi.org/10.21769/BioProtoc.1775},
keywords = {Telomere; G-overhang; In-gel hybridization; Arabidopsis; Telomere analysis},
language = {eng},
issn = {2331-8325},
journal = {Bio-protocol},
title = {Analysis of Telomeric G -overhangs by in- Gel Hybridization},
url = {http://www.bio-protocol.org/e1775},
volume = {6},
year = {2016}
}
TY - JOUR
ID - 1376223
AU - Valuchová, Soňa - Derboven, Elisa - Říha, Karel
PY - 2016
TI - Analysis of Telomeric G -overhangs by in- Gel Hybridization
JF - Bio-protocol
VL - 6
IS - 7
SP - nestránkováno
EP - nestránkováno
PB - Bio-protocol LLC
SN - 23318325
KW - Telomere
KW - G-overhang
KW - In-gel hybridization
KW - Arabidopsis
KW - Telomere analysis
UR - http://www.bio-protocol.org/e1775
L2 - http://www.bio-protocol.org/e1775
N2 - Telomeric DNA in majority of eukaryotes consists of an array of TG-rich tandem repeats. The TG-rich DNA strand is oriented with its 3’ end towards chromosome termini and is usually longer than its complementary CA-rich strand, thus forming 3’ single stranded overhang (G-overhang). G-overhangs arise from incomplete replication of chromosome termini by the lagging strand mechanism and post-replicative nucleolytic processing. The G-overhang is important for telomere protection as it serves as a binding platform for specific proteins and is required for t-loop formation. Hence, structure of telomeric G-overhang is an important indicator of telomere maintenance and functionality. Here we describe a method for analysis of G-overhangs in a model plant Arabidopsis thaliana by in-gel hybridization technique. This method allows relative quantification of the amount of single stranded telomeric DNA. Short telomeric probes are radioactively labeled and hybridized to DNA under non-denaturing conditions to specifically detect ssDNA. Total telomeric DNA can be measured using denaturing conditions in the same gel and this procedure usually follows the non-denaturing in-gel hybridization. Terminal nature of the ssDNA is verified by exonuclease treatment. This technique was originally developed in yeast and now is used as a major tool for G-overhang analysis in variety of organisms ranging from human to plants.
ER -
VALUCHOVÁ, Soňa, Elisa DERBOVEN a Karel ŘÍHA. Analysis of Telomeric G -overhangs by in- Gel Hybridization. \textit{Bio-protocol}. Sunnyvale: Bio-protocol LLC, 2016, roč.~6, č.~7, s.~nestránkováno, 9 s. ISSN~2331-8325. Dostupné z: https://dx.doi.org/10.21769/BioProtoc.1775.
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