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

作者：Sykorova Eva^*; Fojtova Miloslava; Peska Vratislav

来源：Analytical Biochemistry, 2013, 439(1): 8-10.

DOI：10.1016/j.ab.2013.03.034

• BAL-31 nuclease
• Subtelomere
• Interstitial telomere sequence
• Genome sequencing
• Arabidopsis thaliana

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.

• 出版日期2013-8-1