Document 0678
 DOCN  M9650678
 TI    In situ PCR: protocols and applications.
 DT    9605
 AU    Nuovo GJ; Department of Pathology, State University of New York at
       Stony; Brook 11794, USA.
 SO    PCR Methods Appl. 1995 Feb;4(4):S151-67. Unique Identifier : AIDSLINE
       MED/96000051
 AB    Many groups have now published data based on the in situ detection of
       PCR-amplified DNA and cDNA. As with standard in situ hybridization or
       PCR, variables that can affect in situ PCR results include type of
       fixative and time of fixation, protease digestion, and the composition
       of the amplifying solution and oligoprobe cocktail. Investigators new to
       the field of in situ PCR should first try direct incorporation of the
       reporter molecule into paraffin-embedded tissue sections. Although
       nonspecific DNA synthesis is generated under these conditions, one can
       develop the confidence of synthesizing DNA inside the nucleus and
       appreciate the importance of protease digestion time to successful RT in
       situ PCR. It is an arguable statement that the in situ detection of
       PCR-amplified DNA and cDNA will have a very strong impact on many
       diverse fields, such as oncogenesis, embryology, RNA trafficking, and
       detection of viral diseases, as it already has on our understanding of
       the pathogenesis of HIV-1 infection.
 DE    DNA/*ANALYSIS/BLOOD  DNA, Complementary/ANALYSIS  Fixatives
       Histological Techniques  Human  In Situ Hybridization/*METHODS
       Indicators and Reagents  Monocytes/CYTOLOGY/METABOLISM  Oligonucleotide
       Probes  Peptide Peptidohydrolases  Polymerase Chain Reaction/*METHODS
       Proto-Oncogene Proteins/BIOSYNTHESIS  Proto-Oncogenes  RNA-Directed DNA
       Polymerase  Support, Non-U.S. Gov't  JOURNAL ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).