PMID- 12707781
OWN - NLM
STAT- MEDLINE
DCOM- 20040108
LR  - 20181113
IS  - 0009-5915 (Print)
IS  - 0009-5915 (Linking)
VI  - 111
IP  - 7
DP  - 2003 Apr
TI  - Mouse telomere analysis using an optimized primed in situ (PRINS) labeling 
      technique.
PG  - 438-44
AB  - Telomeres are chromosomal elements composed of variable numbers of a TTAGGG 
      repeated DNA sequence required for genomic stability. Telomeric length is 
      correlated with the number of copies of this repeated DNA sequence and is an 
      important property relevant to telomeric function. Recently, it has been 
      demonstrated that the length of the shortest telomere, not average telomeric 
      length, is important for cell viability and chromosomal stability. Consequently, 
      assays permitting assessment of telomeric length are important for the analysis 
      of genomic instability disorders. The length of individual telomeres can be 
      analyzed using the primed in situ (PRINS) labeling reaction, which produces a 
      labeled copy of the telomeric DNA repeats in situ. In this study, we tested 
      different variables to optimize the PRINS reaction to enable it to be applied to 
      the detection of mouse telomeric DNA and the study of telomeric length. The 
      specificity, efficiency and uniformity of staining were evaluated using digital 
      fluorescence microscopy. Labeling efficiency is dependent upon the conditions 
      used to denature the telomeric DNA and reaction duration. Staining uniformity is 
      increased at higher annealing and elongation temperatures as well as when a 
      fluorescently labeled nucleotide is incorporated during the elongation step. Our 
      results also indicate that chromosomal background staining is observed when a 
      fluorochrome-labeled nucleotide is used as opposed to a hapten-labeled 
      nucleotide. From this study, we conclude that an optimized PRINS technique can be 
      reliably employed to analyze mouse telomeres and, compared with the FISH 
      (fluorescence in situ hybridization) technique, presents advantages including 
      greater cost efficiency and reduced processing time. These advantages may 
      encourage wider use of the PRINS technique for quantitative evaluation of the 
      length of individual telomeres in situ.
FAU - Lavoie, Josee
AU  - Lavoie J
AD  - Division de Pathologie, Departement de Biologie Medicale, Universite Laval et 
      Unite de Recherche en Genetique Humaine et Moleculaire, Centre de Recherche de 
      l'Hopital Saint-Francois d'Assise, CHUQ, 10 rue de l'Espinay, Quebec, QC G1L 3L5, 
      Canada.
FAU - Bronsard, Marc
AU  - Bronsard M
FAU - Lebel, Michel
AU  - Lebel M
FAU - Drouin, Regen
AU  - Drouin R
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20030311
PL  - Austria
TA  - Chromosoma
JT  - Chromosoma
JID - 2985138R
SB  - IM
MH  - Animals
MH  - Chromatids/ultrastructure
MH  - Chromosomes/ultrastructure
MH  - Fibroblasts/metabolism
MH  - In Situ Hybridization, Fluorescence
MH  - Mice
MH  - Microscopy, Fluorescence
MH  - Primed In Situ Labeling/*methods
MH  - Repetitive Sequences, Nucleic Acid
MH  - Telomere/*ultrastructure
MH  - Temperature
EDAT- 2003/04/23 05:00
MHDA- 2004/01/09 05:00
CRDT- 2003/04/23 05:00
PHST- 2002/09/09 00:00 [received]
PHST- 2002/10/30 00:00 [revised]
PHST- 2002/11/28 00:00 [accepted]
PHST- 2003/04/23 05:00 [pubmed]
PHST- 2004/01/09 05:00 [medline]
PHST- 2003/04/23 05:00 [entrez]
AID - 10.1007/s00412-002-0225-1 [doi]
PST - ppublish
SO  - Chromosoma. 2003 Apr;111(7):438-44. doi: 10.1007/s00412-002-0225-1. Epub 2003 Mar 
      11.