Molecular and Behavioral Neuroscience Institute, Department of Psychiatry, University of Michigan, Ann Arbor, MI, USA.
Biotechniques. 2012 Sep;53(3):175-80. doi: 10.2144/0000113912.
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.
对离散脑区进行分子分析很复杂，需要在准确性、灵活性和成本之间进行权衡。我们开发了一种灵活且经济有效的方法，即原位杂交（ISH）引导的冷冻基质辅助打孔（IFAP），可从薄至 12μm 的载玻片上的切片中提取纳克量的 DNA。通过使用 ISH 对感兴趣的区域进行定位，将一小滴 M-1 包埋基质施加到冷冻切片上，然后进行冷冻，收集珠子进行核酸纯化，从而对组织进行靶向处理。该方法可定量回收用于 PCR 和 DNA 甲基化分析的 RNA 和 DNA。
