PMID- 17284819
OWN - NLM
STAT- MEDLINE
DCOM- 20070509
LR  - 20070223
IS  - 0916-8451 (Print)
IS  - 0916-8451 (Linking)
VI  - 71
IP  - 2
DP  - 2007 Feb
TI  - Non-denaturing fluorescence in situ hybridization to find replication origins in 
      a specific genome region on the DNA fiber.
PG  - 627-32
AB  - Fluorescence in situ hybridization (FISH) is a useful method of determining the 
      replication timing of specific genomic loci in mammals and of delineating 
      replicon structures on DNA fibers in combination with in vivo replication 
      labeling. In the case of simultaneous detection of a FISH probe and replicated 
      forks, however, the DNA fibers are damaged by the DNA denaturation step for FISH 
      detection, and the resulting fragmented fluorescence signals prevent analysis at 
      high resolution. Here we found that hybridization of the probe to the genomic DNA 
      was possible even under non-denaturing condition, but only at the time its 
      genomic region replicated. Using the method designated non-denaturing FISH, we 
      determined the replication timing of a specific BAC clone and the standard 
      clones, and found that at least one replication origin exists within the genomic 
      region covered by its BAC clone as an example.
FAU - Sugimura, Kazuto
AU  - Sugimura K
AD  - Laboratory of Molecular and Cellular Biology, Graduate School of Bioresources, 
      Mie University, Tsu, Mie, Japan.
FAU - Takebayashi, Shin-ichiro
AU  - Takebayashi S
FAU - Ogata, Shin
AU  - Ogata S
FAU - Taguchi, Hiroshi
AU  - Taguchi H
FAU - Okumura, Katsuzumi
AU  - Okumura K
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20070207
PL  - England
TA  - Biosci Biotechnol Biochem
JT  - Bioscience, biotechnology, and biochemistry
JID - 9205717
RN  - 0 (DNA Probes)
RN  - 9007-49-2 (DNA)
SB  - IM
MH  - Cloning, Molecular
MH  - DNA/*chemistry/*genetics
MH  - DNA Probes
MH  - DNA Replication/physiology
MH  - Genome
MH  - HeLa Cells
MH  - Humans
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Nucleic Acid Denaturation
EDAT- 2007/02/08 09:00
MHDA- 2007/05/10 09:00
CRDT- 2007/02/08 09:00
PHST- 2007/02/08 09:00 [pubmed]
PHST- 2007/05/10 09:00 [medline]
PHST- 2007/02/08 09:00 [entrez]
AID - JST.JSTAGE/bbb/60662 [pii]
AID - 10.1271/bbb.60662 [doi]
PST - ppublish
SO  - Biosci Biotechnol Biochem. 2007 Feb;71(2):627-32. doi: 10.1271/bbb.60662. Epub 
      2007 Feb 7.