Laboratory of Biochemistry and Genetics, National Institute of Diabetes & Digestive and Kidney Diseases, Bethesda, MD, USA.
Methods Mol Biol. 2022;2468:43-49. doi: 10.1007/978-1-0716-2181-3_3.
One of the key attributes that has contributed to the popularity of Caenorhabditis elegans as a model system is its ability to survive freezing. By preserving stocks at ultralow temperature, researchers have been able to generate an unlimited number of strains without the burden of constantly maintaining them. This has facilitated the use of large-scale forward genetic screens and CRISPR-mediated genome editing where large numbers of novel and informative mutants can be generated. Traditionally, C. elegans and other nematodes were frozen using glycerol as a cryoprotectant. While effective, a large majority of animals do not survive a typical freeze-thaw cycle. Here I describe an alternative method based on the popular combination of DMSO and trehalose as a cryoprotectant. This method allows the survival of large numbers of worms and effectively protects most developmental stages including adults.
秀丽隐杆线虫作为一种模型系统广受欢迎的一个关键属性是其能够耐受冷冻。通过将品系保存在超低温下，研究人员能够在无需不断维持的情况下生成无限数量的品系。这促进了大规模正向遗传筛选和 CRISPR 介导的基因组编辑的应用，其中可以生成大量新颖且信息丰富的突变体。传统上，秀丽隐杆线虫和其他线虫使用甘油作为冷冻保护剂进行冷冻。虽然有效，但绝大多数动物无法在典型的冻融循环中存活。在这里，我描述了一种基于二甲亚砜和海藻糖这一流行组合的替代冷冻保护剂方法。这种方法允许大量线虫存活，并有效地保护包括成虫在内的大多数发育阶段。
