利用具有 DNA 水解活性的脱氧核酶生物技术生产单链 DNA。

Biotechnological production of ssDNA with DNA-hydrolyzing deoxyribozymes.

机构信息

Fudan University Shanghai Cancer Center and the Shanghai Key Laboratory of Medical Epigenetics, Institutes of Biomedical Sciences, Shanghai Stomatological Hospital, Fudan University, Shanghai 200433, China.

Center for Medical Research and Innovation, Shanghai Pudong Hospital, Fudan University Pudong Medical Center, Shanghai 201399, China.

出版信息

STAR Protoc. 2021 May 10;2(2):100531. doi: 10.1016/j.xpro.2021.100531. eCollection 2021 Jun 18.

DOI:10.1016/j.xpro.2021.100531

PMID:34027488

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8134756/

Abstract

Preparation of long single-stranded (ss)DNA in large quantities with high efficiency and purity remains a synthetic challenge. Here, we present a protocol for using DNA-hydrolyzing DNA enzymes (deoxyribozymes) for efficient biotechnological production of milligrams of ssDNA with a customizable sequence up to a few kilobases. Our protocol provides a convenient yet economical way to store the sequence information of target ssDNA on phages for selective mass production on demand. For complete details on the use and execution of this protocol, please refer to Jia et al. (2021).

摘要

大量高效且高纯度地制备长单链 DNA 仍然是一个合成难题。在此，我们提供了一种使用 DNA 水解酶（脱氧核酶）的方案，用于高效生物技术生产毫克级至几千碱基对的定制序列的单链 DNA。我们的方案为在噬菌体上存储目标单链 DNA 的序列信息提供了一种方便且经济的方法，可按需进行大规模选择性生产。有关该方案使用和执行的完整详细信息，请参考 Jia 等人（2021 年）。