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Methods in Mammary Gland Biology and Breast Cancer Research pp 223–232Cite as

Application of In Situ PCR to Studies of the Mammary Gland

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Abstract

The ability to detect low numbers of target nucleic acid sequence provides a powerful tool to investigate the molecular events that underlie development and disease in organs such as the mammary gland. Recent development of the polymerase chain reaction in situ (IS-PCR) now enables the intracellular amplification of DNA by PCR and its subsequent localization in preparations of fixed cells and tissue. This chapter outlines the procedure and materials required for the localization of integrated mouse mammary tumor virus proviral DNA in mouse mammary tissue by an indirect IS-PCR method. Also discussed are several aspects critical to the success of this emerging technique, particularly regarding tissue pretreatment and the importance of appropriate controls. In addition, various potential applications of this technique to studies of the mammary gland are considered.

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Abbreviations

(DEPC):

diethylpyrocarbonate

(DTT):

dithiothreitol

(IS-PCR):

in situ PCR

(MMLV):

Moloney murine leukemia virus

(MMTV):

mouse mammary tumor virus

(PBS):

phosphate buffered saline

(PCR):

polymerase chain reaction

(RTPCR):

reverse transcriptase polymerase chain reaction

(rTth):

recombinant Thermus thermophilus DNA polymerase

(SSC):

sodium chloride/sodium citrate

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Author information

Authors and Affiliations

  1. Laboratory of Tumor Immunology and Biology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, 20892, USA

    Russell C. Hovey & Barbara K. Vonderhaar

Authors
  1. Russell C. Hovey
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  2. Barbara K. Vonderhaar
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Editor information

Editors and Affiliations

  1. Department of Pharmacology and Therapeutics, Roswell Park Cancer Institute, Buffalo, New York, 14263, USA

    Margot M. Ip

  2. Roswell Park Cancer Research Institute, Buffalo, New York, USA

    Bonnie B. Asch

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© 2000 Springer Science+Business Media New York

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Hovey, R.C., Vonderhaar, B.K. (2000). Application of In Situ PCR to Studies of the Mammary Gland. In: Ip, M.M., Asch, B.B. (eds) Methods in Mammary Gland Biology and Breast Cancer Research. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-4295-7_20

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  • DOI: https://doi.org/10.1007/978-1-4615-4295-7_20

  • Publisher Name: Springer, Boston, MA

  • Print ISBN: 978-1-4613-6927-1

  • Online ISBN: 978-1-4615-4295-7

  • eBook Packages: Springer Book Archive

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