A polymerase chain reaction-based approach for evaluation of telomere-associated sequences and interstitial telomeric sequences

Telomere minisatellites could be present in both terminal and internal chromosomal regions. We monitored the progress of BAL-31 nuclease digestion on Arabidopsis thaliana genomic DNA prepared by standard isolation techniques to verify its cleavage at terminal and internal genomic regions. A subtelom...

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Bibliographic Details
Published in:Analytical biochemistry 2013-08, Vol.439 (1), p.8-10
Main Authors: Sykorova, Eva, Fojtova, Miloslava, Peska, Vratislav
Format: Article
Language:English
Subjects:
Arabidopsis - genetics
Arabidopsis thaliana
BAL-31 nuclease
Base Sequence
digestion
DNA
DNA, Plant - genetics
Genome sequencing
genomics
Interstitial telomere sequence
isolation techniques
minisatellite repeats
PCR
Polymerase Chain Reaction - methods
quantitative polymerase chain reaction
Subtelomere
Telomere - genetics
telomeres
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Summary:Telomere minisatellites could be present in both terminal and internal chromosomal regions. We monitored the progress of BAL-31 nuclease digestion on Arabidopsis thaliana genomic DNA prepared by standard isolation techniques to verify its cleavage at terminal and internal genomic regions. A subtelomeric position of candidate sequences was validated using conventional polymerase chain reaction (PCR), combining the C-strand-specific telomeric primer with a subtelomeric reverse primer, and confirmed by quantitative PCR (qPCR) using sequence-specific primer pairs on DNA samples after BAL-31 digestion. qPCR amplification showed a gradual decrease in subtelomeric sequence signals, in contrast to interstitial telomeric sequences from pericentromere and control sequences.
ISSN:0003-2697
1096-0309
DOI:10.1016/j.ab.2013.03.034