title: Measurement of telomere length using PNA probe by cytometry.
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.
authors: Clinical Medicine Department, University Sapienza, viale dell'Università, Rome, Italy.
authors: Maurizio
authors: M
authors: Carbonari
authors: Marina
authors: M
authors: Cibati
authors: Nicla
authors: N
authors: Sette
authors: Angela
authors: A
authors: Catizone
authors: Massimo
authors: M
authors: Fiorilli
mesh_terms: D001483:Base Sequence; D002232:Carbocyanines / Q000032:analysis / Q000378:metabolism; D004247:DNA / Q000737:chemistry* / Q000235:genetics; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes / Q000032:analysis / Q000378:metabolism; D005559:Formamides / Q000737:chemistry; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D018613:Microscopy, Confocal; D008969:Molecular Sequence Data; D015341:Nucleic Acid Probes / Q000032:analysis* / Q000138:chemical synthesis; D020135:Peptide Nucleic Acids / Q000032:analysis* / Q000138:chemical synthesis; D011003:Ploidies; D013601:T-Lymphocytes / Q000737:chemistry / Q000473:pathology*; D019098:Telomerase / Q000378:metabolism; D016615:Telomere / Q000737:chemistry* / Q000235:genetics; D014407:Tumor Cells, Cultured
chemical_list: D002232:Carbocyanines; D005456:Fluorescent Dyes; D005559:Formamides; D015341:Nucleic Acid Probes; D020135:Peptide Nucleic Acids; C085321:cyanine dye 5; C031066:formamide; D004247:DNA; D019098:Telomerase