PMID- 8401492
OWN - NLM
STAT- MEDLINE
DCOM- 19931105
LR  - 20190512
IS  - 0964-6906 (Print)
IS  - 0964-6906 (Linking)
VI  - 2
IP  - 8
DP  - 1993 Aug
TI  - Generation of band-specific painting probes from a single microdissected 
      chromosome.
PG  - 1117-21
AB  - We have developed a modified strategy for the generation of regional probes for 
      human chromosomes by microdissection and degenerate oligonucleotide primed PCR. 
      This modification dramatically increases the efficiency of amplification by 
      pretreatment of the dissected chromatin with topoisomerase I (Topo I) before PCR. 
      This protocol has enabled us to construct region-specific probes for fluorescence 
      in situ hybridization (FISH) from a single microdissected chromosome. Results are 
      presented which convincingly demonstrate that this new method generates high 
      intensity region-specific FISH probes, while at the same time significantly 
      decreasing the time-consuming and labor-intensive aspects of microdissection. The 
      reduction of the number of copies required to generate a useful probe also 
      significantly decreases the risk of contamination during the microdissection 
      process. We believe this advance will allow microdissection to be more widely 
      used in the cytogenetic analysis of chromosome rearrangements in both cancer and 
      hereditary diseases. In addition, this method now makes it possible to construct 
      a series of non-overlapping band-specific DNA microclone libraries to provide 
      complete coverage of individual chromosomes for physical mapping.
FAU - Guan, X Y
AU  - Guan XY
AD  - Department of Radiation Oncology, University of Michigan, Ann Arbor 48109-0668.
FAU - Trent, J M
AU  - Trent JM
FAU - Meltzer, P S
AU  - Meltzer PS
LA  - eng
GR  - CA-29476/CA/NCI NIH HHS/United States
GR  - HG-00209/HG/NHGRI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - England
TA  - Hum Mol Genet
JT  - Human molecular genetics
JID - 9208958
RN  - 0 (Chromatin)
RN  - 0 (DNA Primers)
RN  - 0 (DNA Probes)
RN  - EC 5.99.1.2 (DNA Topoisomerases, Type I)
SB  - IM
MH  - Base Sequence
MH  - Chromatin/ultrastructure
MH  - Chromosome Banding
MH  - *Chromosomes, Human/ultrastructure
MH  - DNA Primers
MH  - *DNA Probes
MH  - DNA Topoisomerases, Type I
MH  - Humans
MH  - In Situ Hybridization, Fluorescence
MH  - Karyotyping
MH  - Molecular Sequence Data
MH  - Polymerase Chain Reaction/*methods
EDAT- 1993/08/01 00:00
MHDA- 1993/08/01 00:01
CRDT- 1993/08/01 00:00
PHST- 1993/08/01 00:00 [pubmed]
PHST- 1993/08/01 00:01 [medline]
PHST- 1993/08/01 00:00 [entrez]
AID - 10.1093/hmg/2.8.1117 [doi]
PST - ppublish
SO  - Hum Mol Genet. 1993 Aug;2(8):1117-21. doi: 10.1093/hmg/2.8.1117.