title: ISH-guided freeze-matrix assisted punches: technique for extracting punches from thin slide-mounted tissues for DNA methylation analysis.
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.
authors.affiliation: Molecular and Behavioral Neuroscience Institute, Department of Psychiatry, University of Michigan, Ann Arbor, MI, USA. Merk@RiNA-GmbH.eu
authors.forename: Lawrence S
authors.initials: LS
authors.lastname: Own
authors.forename: Paresh D
authors.initials: PD
authors.lastname: Patel
mesh_terms: D000818:Animals; D001483:Base Sequence; D001923:Brain Chemistry; D004247:DNA / Q000032:analysis* / Q000235:genetics; D019175:DNA Methylation; D017403:In Situ Hybridization / Q000379:methods*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D012680:Sensitivity and Specificity
chemical_list: D004247:DNA