Sequential injection analysis for optimization of molecular biology reactions.
Sequential injection analysis for optimization of molecular biology reactions.
复制标题
用于优化分子生物学反应的顺序注射分析。
DOI：
10.1021/ac103098u
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发表时间：
2011
影响因子：
7.4
通讯作者：
Ellington,AndrewD
中科院分区：
文献类型：
--
作者：
Allen,PeterB;Ellington,AndrewD
为了自动优化复杂的生化和分子生物学反应，我们开发了一个顺序注射分析（SIA）设备，并结合实验设计（DOE）算法。这种硬件和软件的组合自动探索反应的参数空间，并为优化反应条件提供连续的反馈。作为一个例子，我们优化了荧光底物的核酸内切酶消化，并表明优化的反应条件也适用于装置外底物的消化和质粒的消化。序贯技术快速达到优化的反应条件，使用的试剂比批处理（如流体处理机器人并行探索多个反应条件）少。该装置和方法现在应该能够适应更复杂的分子生物学反应，其可变空间相应地更大。
In order to automate the optimization of complex biochemical and molecular biology reactions, we developed a sequential injection analysis (SIA) device and combined this with a design of experiment (DOE) algorithm. This combination of hardware and software automatically explores the parameter space of the reaction and provides continuous feedback for optimizing reaction conditions. As an example, we optimized the endonuclease digest of a fluorogenic substrate and showed that the optimized reaction conditions also applied to the digest of the substrate outside of the device and to the digest of a plasmid. The sequential technique quickly arrived at optimized reaction conditions with less reagent use than a batch process (such as a fluid handling robot exploring multiple reaction conditions in parallel) would have. The device and method should now be amenable to much more complex molecular biology reactions whose variable spaces are correspondingly larger.