PMID- 7508363
OWN - NLM
STAT- MEDLINE
DCOM- 19940311
LR  - 20190830
IS  - 0009-5915 (Print)
IS  - 0009-5915 (Linking)
VI  - 102
IP  - 9
DP  - 1993 Nov
TI  - The development of chromosome-specific composite DNA probes for the mouse and 
      their application to chromosome painting.
PG  - 591-8
AB  - The speed and ease of human cytogenetic analysis has been greatly enhanced by the 
      technique of fluorescence in situ hybridization (FISH). Non-radioactive 
      fluorescently tagged complex DNA probes specific for individual chromosomes can 
      be hybridized to conventionally obtained metaphase chromosome spreads. Several 
      chromosomes may be "painted" concurrently by using combinations of different 
      labeled probes. Surveys of chromosome breakage and rearrangement may be performed 
      very quickly by avoiding the time consuming process of GTG-banding. The 
      application of FISH to mouse cytogenetics would allow large scale molecular 
      toxicology studies to be conducted on the effects of such environmental insults 
      as potential carcinogens, mutagens and radiation. Progress has been hampered, 
      however, as the Mus musculus karyotype consists of 40 acrocentric chromosomes of 
      approximately the same size, making the recognition and separation of individual 
      chromosomes very difficult. We now describe the successful production and 
      application of chromosome-specific composite DNA probes for M. musculus 
      chromosomes 2 and 8. Stable Robertsonian translocated chromosomes were isolated 
      on a flow sorter and their DNA subsequently amplified by degenerate 
      oligonucleotide primer (DOP) PCR. Small pools (300 copies) of each chromosome 
      were denatured at 94 degrees C then annealed with the primer at 30 degrees C for 
      15 cycles. This was followed by 20 cycles at an annealing temperature of 62 
      degrees C. Additional amplification was performed at an annealing temperature of 
      62 degrees C. The chromosome-specific DNA was labeled with biotin 11-dUTP by nick 
      translation and used for FISH. The usefulness of the technique for translocation 
      detection is demonstrated by analyzing chromosome exchanges induced in mice 
      irradiated with 137Cs gamma rays.
FAU - Breneman, J W
AU  - Breneman JW
AD  - Biology and Biotechnology Research Program, Lawrence Livermore National 
      Laboratory, Livermore, CA 94551-9900.
FAU - Ramsey, M J
AU  - Ramsey MJ
FAU - Lee, D A
AU  - Lee DA
FAU - Eveleth, G G
AU  - Eveleth GG
FAU - Minkler, J L
AU  - Minkler JL
FAU - Tucker, J D
AU  - Tucker JD
LA  - eng
GR  - CA55861-01/CA/NCI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - Austria
TA  - Chromosoma
JT  - Chromosoma
JID - 2985138R
RN  - 0 (DNA Probes)
RN  - 9007-49-2 (DNA)
RN  - DVW027E7NL (Chromomycin A3)
RN  - LHQ7J5KV9B (Bisbenzimidazole)
SB  - IM
MH  - Animals
MH  - Base Sequence
MH  - Bisbenzimidazole
MH  - Cells, Cultured
MH  - Chromomycin A3
MH  - *Chromosomes
MH  - DNA/genetics/radiation effects
MH  - DNA Damage
MH  - *DNA Probes
MH  - Female
MH  - Flow Cytometry
MH  - Gamma Rays
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Karyotyping
MH  - Mice/*genetics
MH  - Molecular Sequence Data
MH  - Translocation, Genetic
EDAT- 1993/11/01 00:00
MHDA- 1993/11/01 00:01
CRDT- 1993/11/01 00:00
PHST- 1993/11/01 00:00 [pubmed]
PHST- 1993/11/01 00:01 [medline]
PHST- 1993/11/01 00:00 [entrez]
AID - 10.1007/BF00352306 [doi]
PST - ppublish
SO  - Chromosoma. 1993 Nov;102(9):591-8. doi: 10.1007/BF00352306.