ISH-guided freeze-matrix assisted punches: technique for extracting punches from thin slide-mounted tissues for DNA methylation analysis

BioTechniques

Paresh D. Patel¹  Lawrence S. Own¹ 

[1] 1Molecular and Behavioral Neuroscience Institute, Department of Psychiatry, and Neuroscience Graduate Program, University of Michigan, Ann Arbor, MI, USA;

关键词: DNA methylation;    tissue extraction;    in situ hybridization;    sodium bisulfite sequencing;    tryptophan hydroxylase 2;    TPH2;   

DOI  :  10.2144/0000113912

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown