ISH-Guided Freeze-Matrix Assisted Punches: Technique for Extracting Punches from thin Slide-Mounted Tissues for DNA Methylation Analysis | Zendy

Lawrence S. Own | Zendy; Paresh D. Patel | Zendy

Open Access

ISH-Guided Freeze-Matrix Assisted Punches: Technique for Extracting Punches from thin Slide-Mounted Tissues for DNA Methylation Analysis

Author(s) -

Lawrence S. Own,

Paresh D. Patel

Publication year - 2012

Publication title -

biotechniques

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.617

H-Index - 131

eISSN - 1940-9818

pISSN - 0736-6205

DOI - 10.2144/0000113912

Subject(s) - dna , matrix (chemical analysis) , nucleic acid , dna methylation , in situ hybridization , microbiology and biotechnology , biology , materials science , computational biology , chemistry , chromatography , biochemistry , messenger rna , gene expression , gene

Dissection of discrete brain regions for molecular analysis is complicated by trade-offs between accuracy, flexibility, and costs. We developed a flexible and cost-effective method, in situ hybridization (ISH) guided freeze-matrix assisted punches (IFAP), for extracting nanogram quantities of DNA from slide-mounted sections as thin as 12 µm. Using ISH to localize regions of interest, tissue is targeted by applying a small bead of M-1 embedding matrix onto cryosections, snap-freezing, and collecting the beads for nucleic acid purification. The method quantitatively recovers RNA and DNA usable for PCR and DNA methylation analysis.

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