PMID- 9272175
OWN - NLM
STAT- MEDLINE
DCOM- 19970925
LR  - 20190722
IS  - 0340-6717 (Print)
IS  - 0340-6717 (Linking)
VI  - 100
IP  - 3-4
DP  - 1997 Sep
TI  - Removal of repetitive sequences from FISH probes using PCR-assisted affinity 
      chromatography.
PG  - 472-6
AB  - The vast majority of probes used in fluorescence in situ hybridization (FISH) 
      contain repetitive DNA. This DNA is usually competed out of a hybridization 
      reaction by the addition of an unlabeled blocking agent, Cot-1 DNA. We have 
      successfully removed repetitive DNA from two complex FISH probe sets: a 
      degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR) single 
      human chromosome library and genomic DNA. The procedure involved hybridizing in 
      solution a DOP-PCR-amplifiable probe set with a 50-fold excess of biotin-labeled 
      Cot-1 DNA, and capturing the Cot-1 DNA-containing hybrids using streptavidin 
      magnetic particles, followed by purification and reamplification of the unbound 
      fraction. Probes were checked for depletion of repeats by hybridization to 
      chromosomes without Cot-1 DNA. Results showed hybridization patterns comparable 
      to those achieved with untreated probes hybridized with Cot-1 DNA.
FAU - Craig, J M
AU  - Craig JM
AD  - Institute of Anthropology and Human Genetics, University of Munich, Germany.
FAU - Kraus, J
AU  - Kraus J
FAU - Cremer, T
AU  - Cremer T
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Germany
TA  - Hum Genet
JT  - Human genetics
JID - 7613873
RN  - 9007-49-2 (DNA)
SB  - IM
MH  - Chromatography, Affinity/*methods
MH  - DNA
MH  - Humans
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Male
MH  - Polymerase Chain Reaction/*methods
MH  - *Repetitive Sequences, Nucleic Acid
EDAT- 1997/09/01 00:00
MHDA- 1997/09/01 00:01
CRDT- 1997/09/01 00:00
PHST- 1997/09/01 00:00 [pubmed]
PHST- 1997/09/01 00:01 [medline]
PHST- 1997/09/01 00:00 [entrez]
AID - 10.1007/s004390050536 [doi]
PST - ppublish
SO  - Hum Genet. 1997 Sep;100(3-4):472-6. doi: 10.1007/s004390050536.