Bettscheider Marc, Murgatroyd Chris, Spengler Dietmar
Department of Molecular Neuroendocrinology, Max-Planck-Institute of Psychiatry, Kraepelinstr, 2-10, 80804 Munich, Germany.
BMC Res Notes. 2011 Aug 30;4:314. doi: 10.1186/1756-0500-4-314.
Epigenetic modifications such as DNA methylation play an important role for gene expression and are regulated by developmental and environmental signals. DNA methylation typically occurs in a highly tissue- and cell-specific manner. This raises a severe challenge when studying discrete, small regions of the brain where cellular heterogeneity is high and tissue quantity limited. Because gene expression and methylation are often tightly linked it appears of interest to compare both parameters in the same sample.
We present a refined method for the simultaneous extraction of DNA for bisulfite sequencing and RNA for expression analysis from small mouse brain tissue punches. This method can also be easily adapted for other small tissues or cell populations.
The method described herein results in DNA and RNA of a quantity and quality permitting highly reliable bisulfite analysis and quantitative RT-PCR measurements, respectively.
诸如DNA甲基化等表观遗传修饰在基因表达中起重要作用，并受发育和环境信号调控。DNA甲基化通常以高度组织和细胞特异性的方式发生。当研究大脑中离散的小区域时，这带来了严峻挑战，因为这些区域细胞异质性高且组织量有限。由于基因表达和甲基化通常紧密相连，在同一样本中比较这两个参数似乎很有意义。
我们提出了一种改进方法，可从小鼠脑组织小块中同时提取用于亚硫酸氢盐测序的DNA和用于表达分析的RNA。该方法也可轻松适用于其他小组织或细胞群体。
本文所述方法分别产生了数量和质量足以进行高度可靠的亚硫酸氢盐分析和定量RT-PCR测量的DNA和RNA。
