PMID- 7980939
OWN - NLM
STAT- MEDLINE
DCOM- 19950105
LR  - 20061115
IS  - 0736-6205 (Print)
IS  - 0736-6205 (Linking)
VI  - 17
IP  - 2
DP  - 1994 Aug
TI  - A rapid FISH technique for quantitative microscopy.
PG  - 346-8, 350-3
AB  - Results of quantitative microscopy for fluorescence in situ hybridization (FISH) 
      signals with repetitive DNA probes (pUC 1.77 and D15Z1) are reported. A 
      nonenzymatic hybridization technique was applied using fluorescein-12-dUTP 
      labeled DNA probes and a buffer system not containing any formamide or equivalent 
      chemical denaturing agents. Following thermal denaturation, the renaturation time 
      was reduced to less than 30 min. The number of wash steps was reduced to one. For 
      the pUC 1.77 probe, the major binding sites (chromosome 1) were distinguished 
      from the minor binding sites by means of fluorescence intensity and spot size. 
      The intensity variation of the two brightest FISH spots (major binding sites) in 
      the same metaphase was 19% for 15 min renaturation time and 16% for 30 min 
      renaturation time. For the D15Z1 probe, generally four bright spots were visible 
      and tentatively assigned according to chromosome length and centromere position 
      (chromosomes 15 and 9). The intensity variation of each two homologues in the 
      same metaphase spread showed a coefficient of variation of 47% (15 min) and 22% 
      (30 min) for chromosome 15, and 19% (15 min) and 15% (30 min) for chromosome 9. 
      The results indicate that the applied technique can considerably accelerate the 
      FISH procedure and is suited for quantitative microscopy.
FAU - Haar, F M
AU  - Haar FM
AD  - University of Heidelberg, FRG.
FAU - Durm, M
AU  - Durm M
FAU - Aldinger, K
AU  - Aldinger K
FAU - Celeda, D
AU  - Celeda D
FAU - Hausmann, M
AU  - Hausmann M
FAU - Ludwig, H
AU  - Ludwig H
FAU - Cremer, C
AU  - Cremer C
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Biotechniques
JT  - BioTechniques
JID - 8306785
RN  - 0 (DNA Probes)
SB  - IM
MH  - DNA Probes
MH  - Humans
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Microscopy
EDAT- 1994/08/01 00:00
MHDA- 1994/08/01 00:01
CRDT- 1994/08/01 00:00
PHST- 1994/08/01 00:00 [pubmed]
PHST- 1994/08/01 00:01 [medline]
PHST- 1994/08/01 00:00 [entrez]
PST - ppublish
SO  - Biotechniques. 1994 Aug;17(2):346-8, 350-3.