PMID- 8717051
OWN - NLM
STAT- MEDLINE
DCOM- 19961023
LR  - 20190606
IS  - 1088-9051 (Print)
IS  - 1088-9051 (Linking)
VI  - 5
IP  - 1
DP  - 1995 Aug
TI  - Mechanically stretched chromosomes as targets for high-resolution FISH mapping.
PG  - 13-20
AB  - When used with metaphase chromosomes, fluorescence in situ hybridization (FISH) 
      makes it possible to localize probes to individual chromosome bands and to 
      establish the order of probes separated by > or = 2-3 Mb in dual-color 
      hybridizations. We evaluated the use of mechanically stretched chromosomes as 
      hybridization targets for increased mapping resolution. Mapping resolution was 
      tested by pair-wise hybridizations with probes from the 1p32-p33 region, spanning 
      distances from 20 to approximately 1500 kb. Probes separated by > or = 170 kb 
      could be ordered relative to one another and to the centromere-telomere axis of 
      the chromosome. The advantages of the technique are the simple procedure for 
      preparing the slides, the straightforward interpretation of the results, and the 
      ability to score the predominant order from < 10 stretched chromosomes. However, 
      because of the variability of stretching from one sample to another, the 
      calculation of actual physical distances between probes is not possible. To 
      illustrate the utility of this method, we showed that the gene for receptor 
      tyrosine kinase TIE lies centromeric to COL9A2, RLF, and L-MYC genes at 1p32. The 
      use of mechanically stretched chromosomes provides < or = 10-fold increased 
      mapping resolution as compared with conventional metaphase FISH. Thus, the 
      technique effectively bridges the gap between metaphase mapping and 
      ultra-high-resolution mapping (1-300 kb) techniques, such as the DNA fiber FISH.
FAU - Laan, M
AU  - Laan M
AD  - Department of Clinical Chemistry, University of Helsinki, Finland.
FAU - Kallioniemi, O P
AU  - Kallioniemi OP
FAU - Hellsten, E
AU  - Hellsten E
FAU - Alitalo, K
AU  - Alitalo K
FAU - Peltonen, L
AU  - Peltonen L
FAU - Palotie, A
AU  - Palotie A
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Genome Res
JT  - Genome research
JID - 9518021
RN  - 0 (Hypotonic Solutions)
RN  - 0 (Receptors, Cell Surface)
RN  - EC 2.7.10.1 (Receptor Protein-Tyrosine Kinases)
RN  - EC 2.7.10.1 (Receptors, TIE)
SB  - IM
MH  - Adult
MH  - Centrifugation
MH  - Chromosome Mapping/instrumentation/*methods
MH  - Chromosomes, Human/drug effects/genetics/*ultrastructure
MH  - Chromosomes, Human, Pair 1/drug effects/genetics/ultrastructure
MH  - Cosmids/genetics/ultrastructure
MH  - Humans
MH  - Hypotonic Solutions/pharmacology
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Metaphase
MH  - Receptor Protein-Tyrosine Kinases/genetics
MH  - Receptors, Cell Surface/genetics
MH  - Receptors, TIE
MH  - Sensitivity and Specificity
MH  - Stress, Mechanical
EDAT- 1995/08/01 00:00
MHDA- 1995/08/01 00:01
CRDT- 1995/08/01 00:00
PHST- 1995/08/01 00:00 [pubmed]
PHST- 1995/08/01 00:01 [medline]
PHST- 1995/08/01 00:00 [entrez]
AID - 10.1101/gr.5.1.13 [doi]
PST - ppublish
SO  - Genome Res. 1995 Aug;5(1):13-20. doi: 10.1101/gr.5.1.13.