ISH-guided freeze-matrix assisted punches: technique for extracting punches from thin slide-mounted tissues for DNA methylation analysis. (September 2012)

• Record Type: Journal Article
• Authors: Own, Lawrence S.; Patel, Paresh D.
• Is Part Of: Biotechniques. Volume 53:Number 3(2012)
• Page Start: 175
• Page End: 180
• Publication Date: 2012-09
• Subjects: DNA methylation -- tissue extraction -- in situ hybridization -- sodium bisulfite sequencing -- tryptophan hydroxylase 2 -- TPH2 -- micropunch; Biology, Experimental -- Periodicals; Molecular biology -- Periodicals; Medical technology -- Periodicals; Biology, Experimental; Medical technology; Molecular biology; Clinical Laboratory Techniques -- Periodicals; Research -- Periodicals; Medical Laboratory Science -- Periodicals; Periodicals; Electronic journals; 570
• DOI: 10.2144/0000113912
• Languages: English
• ISSNs: 0736-6205

Abstract

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.