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

Publikace: ANALYTICAL BIOCHEMISTRY 439, 8-10 Autoři: Sykorova, E., Fojtova, M., Peska, V. Rok: 2013

Abstrakt

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. (C) 2013 Elsevier Inc. All rights reserved.