PMID- 8969832
OWN - NLM
STAT- MEDLINE
DCOM- 19970311
LR  - 20180928
IS  - 0736-6205 (Print)
IS  - 0736-6205 (Linking)
VI  - 21
IP  - 6
DP  - 1996 Dec
TI  - High-resolution analysis of DNA replication in released chromatin fibers 
      containing 5-bromodeoxyuridine.
PG  - 1050-4
AB  - A strategy has been devised to physically map replication sites in released 
      chromatin of mammalian cells. When added to the culture medium, 
      5-bromodeoxyuridine (BrdU) is incorporated into replicating DNA, partially 
      replacing thymidine. BrdU pulses as short as one minute can be visualized on 
      preparations of straightened chromatin fibers from protein-extracted nuclei by 
      means of monoclonal anti-BrdU antibody. Short BrdU pulses (< 10 min) appear as 
      strings of fluorescent signals that are 50-300 kb in length. This corresponds to 
      the estimated size of individual replication units. Pulse chase experiments 
      reveal that replicating DNA is tightly associated with the residual nuclear 
      matrix, whereas newly replicated DNA is positioned on the released loop chromatin 
      of nuclear halo preparations. Simultaneous fluorescence in situ hybridization 
      (FISH) on BrdU-substituted released chromatin fibers suggests that replication 
      can initiate at multiple sites anywhere within an alpha-satellite array of 
      several Mb.
FAU - Haaf, T
AU  - Haaf T
AD  - Max-Planck-Institute of Molecular Genetics, Berlin, Germany. 
      haaf@mping-berlin-dahlem.mpg.de
LA  - eng
PT  - Technical Report
PL  - England
TA  - Biotechniques
JT  - BioTechniques
JID - 8306785
RN  - 0 (Chromatin)
RN  - 9007-49-2 (DNA)
RN  - G34N38R2N1 (Bromodeoxyuridine)
SB  - IM
MH  - Bromodeoxyuridine/*metabolism
MH  - Cells, Cultured
MH  - Chromatin/*metabolism
MH  - DNA/*analysis
MH  - DNA Replication/*genetics
MH  - Fibroblasts
MH  - Fluorescent Antibody Technique, Indirect
MH  - Humans
MH  - Image Processing, Computer-Assisted
EDAT- 1996/12/01 00:00
MHDA- 1996/12/01 00:01
CRDT- 1996/12/01 00:00
PHST- 1996/12/01 00:00 [pubmed]
PHST- 1996/12/01 00:01 [medline]
PHST- 1996/12/01 00:00 [entrez]
AID - 10.2144/96216st01 [doi]
PST - ppublish
SO  - Biotechniques. 1996 Dec;21(6):1050-4. doi: 10.2144/96216st01.