利用磁珠从水稻营养组织中提取总 RNA。
Total RNA Extraction from Rice Vegetative Tissues Using Magnetic Beads.
机构信息
Department of Agrobiology and Bioresources, Faculty of Agriculture, Utsunomiya University, Utsunomiya, Tochigi, Japan.
Institute of Agrobiological Sciences, National Agriculture and Food Research Organization, Tsukuba, Ibaraki, Japan.
出版信息
Methods Mol Biol. 2025;2869:7-13. doi: 10.1007/978-1-0716-4204-7_2.
Ribonucleic acid (RNA) extraction is the first critical step in gene expression analysis. In this chapter, we describe a high-throughput RNA extraction method using guanidine thiocyanate and isopropyl alcohol (HighGI). The use of carboxyl-coated paramagnetic beads, instead of silica membrane columns, enables semi-automation using a liquid handling system and high-throughput RNA extraction for large-scale transcriptome studies. Homemade mixes of paramagnetic beads and buffers make HighGI inexpensive. In addition, HighGI-extracted RNA retains low molecular weight RNA molecules less than 200 bp, which is typically lost in commercial column-based kits.
RNA 提取是基因表达分析的关键步骤之一。在本章中，我们描述了一种使用异硫氰酸胍和异丙醇（HighGI）的高通量 RNA 提取方法。使用羧基涂层的超顺磁珠代替硅胶膜柱，可使用液体处理系统进行半自动化，并可进行大规模转录组研究的高通量 RNA 提取。超顺磁珠和缓冲液的自制混合物使 HighGI 价格低廉。此外，HighGI 提取的 RNA 保留了低分子量 RNA 分子，小于 200 bp，这通常在商业柱基试剂盒中丢失。
Zotero 插件