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Unlocking the Full Potential of Cas12a: Exploring the Effects of Substrate and Reaction Conditions on Trans-Cleavage Activity
Analytical Chemistry ( IF 6.7 ) Pub Date : 2023-07-01 , DOI: 10.1021/acs.analchem.3c01307 Jie Xu 1 , Zhujun Liu 1 , Zhen Zhang 1 , Tongbo Wu 1
Analytical Chemistry ( IF 6.7 ) Pub Date : 2023-07-01 , DOI: 10.1021/acs.analchem.3c01307 Jie Xu 1 , Zhujun Liu 1 , Zhen Zhang 1 , Tongbo Wu 1
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The trans-cleavage activity of Cas12a has been widely used with various applications. Here, we report that the trans-cleavage activity of Cas12a can be significantly affected by the fluorescent probe length and reaction buffer. The optimal probe length for Cas12a is found to be 15 nucleotides, and the optimal buffer is NEBuffer 4. Compared to the popularly used reaction conditions, the activity of Cas12a is improved by about 50-fold. In addition, the detection limit of Cas12a for DNA targets has been reduced by nearly three orders of magnitude. Our method provides a powerful tool for Cas12a trans-cleavage activity applications.
中文翻译：
释放 Cas12a 的全部潜力：探索底物和反应条件对反式裂解活性的影响
Cas12a 的反式裂解活性已广泛应用于各种应用。在这里，我们报告 Cas12a 的反式切割活性会受到荧光探针长度和反应缓冲液的显着影响。Cas12a的最佳探针长度为15个核苷酸，最佳缓冲液为NEBuffer 4。与常用的反应条件相比，Cas12a的活性提高了约50倍。此外，Cas12a对DNA靶标的检测限降低了近三个数量级。我们的方法为 Cas12a 反式切割活性应用提供了强大的工具。
更新日期：2023-07-01
中文翻译：
释放 Cas12a 的全部潜力：探索底物和反应条件对反式裂解活性的影响
Cas12a 的反式裂解活性已广泛应用于各种应用。在这里，我们报告 Cas12a 的反式切割活性会受到荧光探针长度和反应缓冲液的显着影响。Cas12a的最佳探针长度为15个核苷酸，最佳缓冲液为NEBuffer 4。与常用的反应条件相比，Cas12a的活性提高了约50倍。此外，Cas12a对DNA靶标的检测限降低了近三个数量级。我们的方法为 Cas12a 反式切割活性应用提供了强大的工具。
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