PMID- 1313061
OWN - NLM
STAT- MEDLINE
DCOM- 19920427
LR  - 20170214
IS  - 0022-1554 (Print)
IS  - 0022-1554 (Linking)
VI  - 40
IP  - 3
DP  - 1992 Mar
TI  - Intracellular amplification of proviral DNA in tissue sections using the 
      polymerase chain reaction.
PG  - 333-41
AB  - We developed a new method to amplify cell DNA in situ using the polymerase chain 
      reaction (PCR). Proviral sequences of mouse mammary tumor virus (MMTV) contained 
      in cultured cells and tissue sections were amplified intracellularly using a 
      thermal cycler. Two techniques were employed to maintain the localization of the 
      amplified DNA. First, complementary tails at the 5' ends of the oligonucleotide 
      primers resulted in the synthesis of high molecular weight concatamers containing 
      the target sequences. Second, the PCR was carried out in a thin film of agarose 
      solidified over the tissue sections. The specifically amplified and localized DNA 
      was then detected by in situ hybridization (ISH). Our results demonstrate that 
      (a) DNA in tissue sections can serve as the target for the polymerase chain 
      reaction in situ, (b) cell morphology is maintained, and (c) a target of 167 BP 
      can be specifically detected in individual cells. This technique should be 
      generally applicable to amplifying cellular DNA targets in tissue sections for 
      detection in situ.
FAU - Chiu, K P
AU  - Chiu KP
AD  - Department of Medicine, School of Medicine, University of California, Davis 
      95817.
FAU - Cohen, S H
AU  - Cohen SH
FAU - Morris, D W
AU  - Morris DW
FAU - Jordan, G W
AU  - Jordan GW
LA  - eng
GR  - R01-DK34239/DK/NIDDK NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - J Histochem Cytochem
JT  - The journal of histochemistry and cytochemistry : official journal of the 
      Histochemistry Society
JID - 9815334
RN  - 0 (DNA, Viral)
RN  - 0 (Oligodeoxyribonucleotides)
SB  - IM
MH  - Animals
MH  - Base Sequence
MH  - Cell Line, Transformed
MH  - DNA, Viral/*analysis/genetics
MH  - Fibroblasts/microbiology/ultrastructure
MH  - Mammary Neoplasms, Experimental/*pathology/ultrastructure
MH  - Mammary Tumor Virus, Mouse/genetics/*isolation & purification
MH  - Mice
MH  - Molecular Sequence Data
MH  - Oligodeoxyribonucleotides
MH  - Polymerase Chain Reaction/*methods
MH  - Proviruses/genetics/*isolation & purification
MH  - Repetitive Sequences, Nucleic Acid
EDAT- 1992/03/01 00:00
MHDA- 1992/03/01 00:01
CRDT- 1992/03/01 00:00
PHST- 1992/03/01 00:00 [pubmed]
PHST- 1992/03/01 00:01 [medline]
PHST- 1992/03/01 00:00 [entrez]
AID - 10.1177/40.3.1313061 [doi]
PST - ppublish
SO  - J Histochem Cytochem. 1992 Mar;40(3):333-41. doi: 10.1177/40.3.1313061.