Boothby Thomas C
Department of Chemistry, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599
Cold Spring Harb Protoc. 2018 Nov 1;2018(11):2018/11/pdb.prot102376. doi: 10.1101/pdb.prot102376.
Purification of high-quality total RNA from specimens is essential for many molecular techniques. In tardigrades (water bears), disruption of the cuticle is an important step in obtaining a good yield that is representative of all tissues of the animal. As with all single-stranded RNA methods, sterile technique, proper storage conditions, and handling are required for maintaining the quality and integrity of the material. This procedure will result in high-quality total RNA suitable for downstream applications such as cDNA synthesis, reverse transcriptase-polymerase chain reaction (RT-PCR), RNAseq library generation, and northern blots.
从样本中纯化高质量的总RNA对于许多分子技术至关重要。在缓步动物(水熊虫)中,破坏角质层是获得代表动物所有组织的良好产量的重要步骤。与所有单链RNA方法一样,需要无菌技术、适当的储存条件和处理方式来维持材料的质量和完整性。此程序将产生适用于下游应用(如cDNA合成、逆转录聚合酶链反应(RT-PCR)、RNA测序文库生成和Northern印迹)的高质量总RNA。