Li Jun-Hui, Tang Chuan-Hong, Song Chun-Yan, Chen Ming-Jie, Feng Zhi-Yong, Pan Ying-Jie
College of Life Sciences, Nanjing Agricultural University, Nanjing, Jiangsu, 210095, PR China.
Biotechnol Lett. 2006 Aug;28(15):1193-7. doi: 10.1007/s10529-006-9074-y. Epub 2006 Jun 24.
A rapid, inexpensive and reliable method for total RNA extraction from fruiting bodies of Lentinula edodes containing large quantities of polysaccharides and secondary metabolites is described. An initial extraction step using saturated NaCl solution facilitates the separation of nucleic acids from contaminants and, after further extraction with organic solvents and precipitation with 2-propanol, total RNA of high purity and suitable for applications such as cDNA synthesis, RT-PCR and Northern blot hybridization was obtained. The procedure may also have wider applicability for total RNA extraction from the tissues of other mushrooms.
本文描述了一种从含有大量多糖和次生代谢产物的香菇子实体中快速、廉价且可靠地提取总RNA的方法。使用饱和NaCl溶液的初始提取步骤有助于将核酸与污染物分离,在用有机溶剂进一步提取并用2-丙醇沉淀后,获得了高纯度且适用于cDNA合成、RT-PCR和Northern印迹杂交等应用的总RNA。该方法可能也更广泛适用于从其他蘑菇组织中提取总RNA。