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In Situ Hybridization Freeze-Assisted Punches (IFAP): Technique for Liquid-Based Tissue Extraction from Thin Slide-Mounted Sections for DNA Methylation Analysis

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In Situ Hybridization Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1211))

Abstract

In situ hybridization-assisted punches (IFAP) are a low-cost method for extracting tissue from frozen slide-mounted sections as thin as 12 μm. The method synergizes well with standard histological workflows and uses in situ hybridization to target corresponding slide-mounted cryosections that contain the region of interest. Liquid beads of M-1 embedding matrix are applied and snap frozen, binding the matrix to the underlying tissue. Bead-tissue complexes are removed and DNA extracted using a high-salt method. IFAP-extracted DNA is suitable for downstream DNA methylation analysis.

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References

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Acknowledgements

We would like to thank Delia Vazquez, Juan Lopez, Robert Thompson, and Robert Denver for their input. This work was supported by a grant from the Pritzker Neuropsychiatric Disorders Research Consortium, which is supported by the Pritzker Neuropsychiatric Disorders Research Fund L.L.C. and a NARSAD Young Investigator Grant to P.D.P. (N008728).

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Authors and Affiliations

  1. Molecular and Behavioral Neuroscience Institute, University of Michigan, 500 South State Street, Ann Arbor, MI, 48109, USA

    Lawrence S. Own & Paresh D. Patel

  2. Neuroscience Graduate Program, University of Michigan, 4137 Undergraduate Science Building, 204 Washetenaw Avenue, Ann Arbor, MI, 48109, USA

    Lawrence S. Own & Paresh D. Patel

  3. Molecular and Behavioral Neuroscience Institute, University of Michigan, 205 Zina Pitcher Place, Ann Arbor, MI, 48109, USA

    Paresh D. Patel

  4. Department of Psychiatry, University of Michigan, 1500 E. Medical Center Drive, Ann Arbor, MI, 48109, USA

    Paresh D. Patel

  5. Molecular and Behavioral Neuroscience Institute, University of Michigan, 205 Zina Pitcher Place, Biomedical Science Research Building, Room 5698, Ann Arbor, MI, 48109, USA

    Lawrence S. Own

  6. Engineering, Voxa, 1610 N. 170th Street, Shoreline, WA, 98133, USA

    Lawrence S. Own

Authors
  1. Lawrence S. Own
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  2. Paresh D. Patel
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Corresponding author

Correspondence to Lawrence S. Own .

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Editors and Affiliations

  1. Molecular Histology, Bioneer A/S, Horsholm, Denmark

    Boye Schnack Nielsen

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Own, L.S., Patel, P.D. (2014). In Situ Hybridization Freeze-Assisted Punches (IFAP): Technique for Liquid-Based Tissue Extraction from Thin Slide-Mounted Sections for DNA Methylation Analysis. In: Nielsen, B. (eds) In Situ Hybridization Protocols. Methods in Molecular Biology, vol 1211. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1459-3_19

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  • DOI: https://doi.org/10.1007/978-1-4939-1459-3_19

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1458-6

  • Online ISBN: 978-1-4939-1459-3

  • eBook Packages: Springer Protocols

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