Carbonari Maurizio, Cibati Marina, Sette Nicla, Catizone Angela, Fiorilli Massimo
Clinical Medicine Department, University Sapienza, viale dell'Università, Rome, Italy.
Methods Cell Biol. 2011;103:189-202. doi: 10.1016/B978-0-12-385493-3.00008-5.
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.
肽核酸（PNA）探针可与在热甲酰胺中透化的细胞中的变性端粒序列杂交。在已报道的实验方案中，杂交是在高甲酰胺浓度的溶液中进行的，以避免DNA复性，因为DNA复性会妨碍寡聚PNA探针与特定序列的结合。我们推测，局限于核微体积中的端粒DNA在热甲酰胺变性后无法正确复性。因此，为了改善探针与靶序列之间的杂交条件，在完全去除甲酰胺后向样品中添加探针可能是可行的。
