Schwartz Katja, Sherlock Gavin
Department of Genetics, Stanford University Medical School, Stanford, California 94305-5120.
Cold Spring Harb Protoc. 2016 Oct 3;2016(10):2016/10/pdb.prot088930. doi: 10.1101/pdb.prot088930.
This protocol provides a detailed description of how to prepare a DNA sequencing library from yeast genomic DNA for use with the Illumina sequencing platform. This method does not require purchase of Illumina kits for library preparation but instead employs specific reagents purchased largely from New England BioLabs, which significantly reduces the cost of library preparation. Although we assume here that users intend to generate libraries with ∼400-bp insert sizes for paired-end sequencing, it is relatively straightforward to modify the shearing and size selection steps for longer or shorter inserts.
本方案详细描述了如何从酵母基因组DNA制备DNA测序文库,以用于Illumina测序平台。此方法不需要购买Illumina文库制备试剂盒,而是使用主要从新英格兰生物实验室购买的特定试剂,这显著降低了文库制备成本。尽管我们在此假设用户打算生成用于双端测序的插入片段大小约为400bp的文库,但对于更长或更短的插入片段,修改剪切和大小选择步骤相对简单。