PMID- 19108707
OWN - NLM
STAT- PubMed-not-MEDLINE
DCOM- 20110714
LR  - 20240316
IS  - 1755-8166 (Electronic)
IS  - 1755-8166 (Linking)
VI  - 1
DP  - 2008 Dec 23
TI  - Validation of DNA probes for molecular cytogenetics by mapping onto immobilized 
      circular DNA.
PG  - 28
LID - 10.1186/1755-8166-1-28 [doi]
AB  - BACKGROUND: Fluorescence in situ hybridization (FISH) is a sensitive and rapid 
      procedure to detect gene rearrangements in tumor cells using non-isotopically 
      labeled DNA probes. Large insert recombinant DNA clones such as bacterial 
      artificial chromosome (BAC) or P1/PAC clones have established themselves in 
      recent years as preferred starting material for probe preparations due to their 
      low rates of chimerism and ease of use. However, when developing probes for the 
      quantitative analysis of rearrangements involving genomic intervals of less than 
      100 kb, careful probe selection and characterization are of paramount importance. 
      RESULTS: We describe a sensitive approach to quality control probe clones 
      suspected of carrying deletions or for measuring clone overlap with near kilobase 
      resolution. The method takes advantage of the fact that P1/PAC/BAC's can be 
      isolated as circular DNA molecules, stretched out on glass slides and fine-mapped 
      by multicolor hybridization with smaller probe molecules. Two examples 
      demonstrate the application of this technique: mapping of a gene-specific ~6 kb 
      plasmid onto an unusually small, ~55 kb circular P1 molecule and the 
      determination of the extent of overlap between P1 molecules homologous to the 
      human NF-kappaB2 locus. CONCLUSION: The relatively simple method presented here 
      does not require specialized equipment and may thus find widespread applications 
      in DNA probe preparation and characterization, the assembly of physical maps for 
      model organisms or in studies on gene rearrangements.
FAU - Greulich-Bode, Karin M
AU  - Greulich-Bode KM
AD  - Division Genetics of Skin Carcinogenesis, German Cancer Research Center (DKFZ), 
      Heidelberg, Germany. k.greulich@dkfz-heidelberg.de.
FAU - Wang, Mei
AU  - Wang M
FAU - Rhein, Andreas P
AU  - Rhein AP
FAU - Weier, Jingly F
AU  - Weier JF
FAU - Weier, Heinz-Ulli G
AU  - Weier HU
LA  - eng
GR  - R33 CA088258-03/CA/NCI NIH HHS/United States
GR  - R44 HD044313-01/HD/NICHD NIH HHS/United States
GR  - R21 CA123370/CA/NCI NIH HHS/United States
GR  - R21 CA123370-01A1/CA/NCI NIH HHS/United States
GR  - R44 HD044313/HD/NICHD NIH HHS/United States
PT  - Journal Article
DEP - 20081223
PL  - England
TA  - Mol Cytogenet
JT  - Molecular cytogenetics
JID - 101317942
PMC - PMC2630919
EDAT- 2008/12/26 09:00
MHDA- 2008/12/26 09:01
PMCR- 2008/12/23
CRDT- 2008/12/26 09:00
PHST- 2008/11/11 00:00 [received]
PHST- 2008/12/23 00:00 [accepted]
PHST- 2008/12/26 09:00 [entrez]
PHST- 2008/12/26 09:00 [pubmed]
PHST- 2008/12/26 09:01 [medline]
PHST- 2008/12/23 00:00 [pmc-release]
AID - 1755-8166-1-28 [pii]
AID - 10.1186/1755-8166-1-28 [doi]
PST - epublish
SO  - Mol Cytogenet. 2008 Dec 23;1:28. doi: 10.1186/1755-8166-1-28.