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Measurement of telomere length using PNA probe by cytometry.
- Source :
-
Methods in cell biology [Methods Cell Biol] 2011; Vol. 103, pp. 189-202. - Publication Year :
- 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.<br /> (Copyright © 2011 Elsevier Inc. All rights reserved.)
- Subjects :
- Base Sequence
Carbocyanines analysis
Carbocyanines metabolism
DNA genetics
Fluorescent Dyes analysis
Fluorescent Dyes metabolism
Formamides chemistry
Humans
Microscopy, Confocal
Molecular Sequence Data
Nucleic Acid Probes chemical synthesis
Peptide Nucleic Acids chemical synthesis
Ploidies
T-Lymphocytes chemistry
Telomerase metabolism
Telomere genetics
Tumor Cells, Cultured
DNA chemistry
Flow Cytometry methods
In Situ Hybridization, Fluorescence methods
Nucleic Acid Probes analysis
Peptide Nucleic Acids analysis
T-Lymphocytes pathology
Telomere chemistry
Subjects
Details
- Language :
- English
- ISSN :
- 0091-679X
- Volume :
- 103
- Database :
- MEDLINE
- Journal :
- Methods in cell biology
- Publication Type :
- Academic Journal
- Accession number :
- 21722804
- Full Text :
- https://doi.org/10.1016/B978-0-12-385493-3.00008-5