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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

  • Lawrence S. OwnEmail author
  • Paresh D. Patel
Protocol
Part of the Methods in Molecular Biology book series (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.

Key words

IFAP Micropunch Tissue extraction In situ hybridization DNA methylation Histology 

Notes

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).

References

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  1. 1.Molecular and Behavioral Neuroscience InstituteUniversity of MichiganAnn ArborUSA
  2. 2.Neuroscience Graduate ProgramUniversity of MichiganAnn ArborUSA
  3. 3.Molecular and Behavioral Neuroscience InstituteUniversity of MichiganAnn ArborUSA
  4. 4.Department of PsychiatryUniversity of MichiganAnn ArborUSA
  5. 5.Molecular and Behavioral Neuroscience InstituteUniversity of MichiganAnn ArborUSA
  6. 6.EngineeringVoxaShorelineUSA

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