PMID- 11180596
OWN - NLM
STAT- MEDLINE
DCOM- 20010802
LR  - 20061115
IS  - 1098-1004 (Electronic)
IS  - 1059-7794 (Linking)
VI  - 17
IP  - 2
DP  - 2001 Feb
TI  - A new technique for cyclic in situ amplification and a case report about 
      amplification of a single copy gene sequence in human metaphase chromosomes 
      through PCR-PRINS.
PG  - 131-40
AB  - Since the introduction of PRimed IN Situ labeling (PRINS) as a rapid and 
      extremely sensitive alternative method to conventional fluorescence in situ 
      hybridization (FISH), its application in clinical cytogenetics has been limited 
      to the detection of highly repeated sequences, such as centromeric and telomeric 
      regions. In the original PRINS method, unlabeled oligonucleotide probes are 
      annealed to their repeated complementary target sequences in fixed human 
      metaphase chromosomes on a slide. The probes serve as primers for subsequent in 
      situ chain elongation with Taq DNA polymerase and labeled nucleotides. In 
      contrast to conventional PCR, cyclic in situ amplification of the chromosomal 
      target DNA with paired primers remained both difficult and strictly limited to 
      highly repeated sequences, since the maintenance of constant reaction conditions 
      on the slide during temperature and pressure shifts presents a major problem. We 
      developed a new system for in situ PCR that allows the amplification of target 
      sequences analogous to PCR in the test tube. We applied this method successfully 
      for the detection of highly repeated sequences, for the detection of low copy 
      repeats, and in one case, for the detection of a single-copy DNA sequence. The 
      significance of this development for further in situ PCR applications will be 
      discussed.
CI  - Copyright 2001 Wiley-Liss, Inc.
FAU - Harrer, T
AU  - Harrer T
AD  - Department of Human Genetics, Medical University of Lubeck, Lubeck, Germany. 
      mennicke@medinf.mu-luebeck.de
FAU - Schwinger, E
AU  - Schwinger E
FAU - Mennicke, K
AU  - Mennicke K
LA  - eng
PT  - Case Reports
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Hum Mutat
JT  - Human mutation
JID - 9215429
RN  - 0 (Apolipoproteins A)
RN  - 0 (DNA Primers)
RN  - 0 (Dystrophin)
SB  - IM
MH  - Apolipoproteins A/genetics
MH  - Chromosomes, Human/*genetics
MH  - DNA Primers
MH  - Dystrophin/genetics
MH  - Female
MH  - Gene Dosage
MH  - Humans
MH  - Male
MH  - Metaphase/*genetics
MH  - Muscular Dystrophies/genetics
MH  - Polymerase Chain Reaction
MH  - Primed In Situ Labeling/*methods
MH  - Repetitive Sequences, Nucleic Acid/genetics
EDAT- 2001/02/17 11:00
MHDA- 2001/08/03 10:01
CRDT- 2001/02/17 11:00
PHST- 2001/02/17 11:00 [pubmed]
PHST- 2001/08/03 10:01 [medline]
PHST- 2001/02/17 11:00 [entrez]
AID - 10.1002/1098-1004(200102)17:2<131::AID-HUMU5>3.0.CO;2-7 [pii]
AID - 10.1002/1098-1004(200102)17:2<131::AID-HUMU5>3.0.CO;2-7 [doi]
PST - ppublish
SO  - Hum Mutat. 2001 Feb;17(2):131-40. doi: 
      10.1002/1098-1004(200102)17:2<131::AID-HUMU5>3.0.CO;2-7.