一个高通量协议,用于去除长单链DNA和含有复杂序列的基池的复杂序列
Yanyan Wang1, Senxin Zhang2, Xiaoqi Zheng3
1Shanghai Institute of Immunology, State Key Laboratory of Oncogenes and Related Genes, Department of Immunology and Microbiology, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
STAR protocols
|September 24, 2023
概括
这项研究引入了一种高通量方法,用于使用cytidine deaminases,如激活诱导deaminase (AID) 进行DNA去胺. 该协议能够对DNA序列中的突变概况进行详细分析,这对于免疫和基因组稳定性研究至关重要.
科学领域:
- 分子生物学分子生物学
- 遗传学 遗传学 是一个
- 免疫学 免疫学 免疫学
背景情况:
- 丁脱氨酶是关键的DNA突变因子,参与免疫反应和维持基因组稳定.
- 了解它们的活动对于各种生物过程至关重要.
研究的目的:
- 开发一种高通量协议,用于长单链DNA或Oligo池的DNA除.
- 为了能够详细分析复杂DNA序列中的突变概况.
主要方法:
- 该协议包括制备激活诱导脱氨酶 (AID) 酶和单链DNA (ssDNA) 基质.
- 它包括除反应的步骤, uracil 友好的放大,以及随后的数据分析.
主要成果:
- 开发的试验允许确定内在突变配置文件.
- 它可以应用于单个抗体基因或选定的基因组DNA区域的池.
结论:
- 这种高通量除毒协议为研究DNA突变因子活动提供了强大的方法.
- 它对免疫力和基因组稳定性的研究有重大影响.
相关概念视频
DNA Isolation
39.3K
DNA isolation protocols can be fast and straightforward or complex and time-consuming depending on the type and quality of DNA required for further processing. For example, plasmid DNA extraction is a bit more complicated than genomic DNA extraction because of the need for an appropriate lysis method to separate plasmid DNA from gDNA during isolation. However, for specific applications, such as long-range DNA sequencing that require a good yield of high- quality DNA samples, we need to follow...
39.3K
Sanger Sequencing
754.7K
DNA sequencing is a fundamental technique that is routinely used in the biological sciences. This method can be applied to a range of questions at different scales - from the sequencing of a cloned DNA fragment or the study of a mutation in a gene up to whole-genome sequencing. However, despite the widespread use of sequencing today, it was not until 1977 that Fredrick Sanger and his collaborators developed the chain-termination method to decode DNA sequences. It relies on the separation of a...
754.7K


