Peptide nucleic acid (PNA) probes hybridize to denatured telomeric sequences in cells permeabilized in hot formamide. In reported protocols, the hybridization was conducted in solutions with high formamide concentrations to avoid the DNA renaturation that can hamper binding of the oligo-PNA probe to specific sequences. We postulated that telomeric DNA, confined in the nuclear microvolume, is not able to properly renature after hot formamide denaturation. Therefore, to improve hybridization conditions between the probe and the target sequences, it might be possible to add probe to sample after the complete removal of formamide.
Measurement of Telomere Length Using PNA Probe by Cytometry / Carbonari, Maurizio; Marina, Cibati; Nicla, Sette; Catizone, Angiolina; Fiorilli, Massimo. - In: METHODS IN CELL BIOLOGY. - ISSN 0091-679X. - 103:(2011), pp. 189-202. [10.1016/b978-0-12-385493-3.00008-5]
Measurement of Telomere Length Using PNA Probe by Cytometry
CARBONARI, Maurizio;CATIZONE, Angiolina;FIORILLI, Massimo
2011
Abstract
Peptide nucleic acid (PNA) probes hybridize to denatured telomeric sequences in cells permeabilized in hot formamide. In reported protocols, the hybridization was conducted in solutions with high formamide concentrations to avoid the DNA renaturation that can hamper binding of the oligo-PNA probe to specific sequences. We postulated that telomeric DNA, confined in the nuclear microvolume, is not able to properly renature after hot formamide denaturation. Therefore, to improve hybridization conditions between the probe and the target sequences, it might be possible to add probe to sample after the complete removal of formamide.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
