从低成本的寡核酸混合物中高度并行构建DNA,使用数据优化的组装设计和金门
Sean Lund1, Vladimir Potapov1, Sean R Johnson1
1Research Department, New England Biolabs, Ipswich, Massachusetts 01938, United States.
ACS synthetic biology
|February 20, 2024
概括
我们使用数据优化组装设计 (DAD) 和金门组装开发了一种新的基因合成方法. 这种方法迅速产生了编码子优化的基因,使得序列的构建比传统方法更广泛.
科学领域:
- 合成生物学 合成生物学
- 分子生物学分子生物学
- 基因合成 基因合成
背景情况:
- 商业基因合成通常依赖于基于同质性的克隆.
- 现有的方法,如聚合酶循环组合,在序列宽度方面存在局限性.
研究的目的:
- 应用数据优化组装设计 (DAD) 以实现高效的基因合成.
- 利用金门组合在各种矢量中构建编码子优化的基因.
主要方法:
- 从寡核酸池中合成的基因分为三个步骤:部分放大,金门组装和转换.
- 数据优化装配设计 (DAD) 指导了装配过程.
- 用T4DNA结合酶来获得高结合效率.
主要成果:
- 数以百计的编码子优化基因被快速合成,在短短4天内得到了序列确认的分离物.
- 该方法在构成和可诱导向量方面都表现出了效率.
- 该方法预计将支持比基于同质学的方法更广泛的序列范围.
结论:
- DAD与金门组合相结合,为基因合成提供了快速有效的方法.
- 这种技术扩大了构建多种基因序列的可能性.
- 该方法提供了传统基因合成方法的替代方案,具有改进的能力.
相关概念视频
Genome Annotation and Assembly
18.8K
The genome refers to all of the genetic material in an organism. It can range from a few million base pairs in microbial cells to several billion base pairs in many eukaryotic organisms. Genome assembly refers to the process of taking the DNA sequencing data and putting it all back together in a correct order to create a close representation of the original genome. This is followed by the identification of functional elements on the newly assembled genome, a process called genome annotation.
18.8K
Sanger Sequencing
754.3K
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...
754.3K
Next-generation Sequencing
88.8K
The first human genome sequencing project cost $2.7 billion and was declared complete in 2003, after 15 years of international cooperation and collaboration between several research teams and funding agencies. Today, with the advent of next-generation sequencing technologies, the cost and time of sequencing a human genome have dropped over 100 fold.
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features....
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features....
88.8K
DNA Isolation
39.1K
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...
39.1K


