Biomimetic Systems, Max Planck Institute of Biochemistry, Martinsried, Germany.
Faculty of Chemistry and Chemical Biology, TU Dortmund University, Dortmund, Germany.
Methods Mol Biol. 2022;2439:27-44. doi: 10.1007/978-1-0716-2047-2_3.
The ability to produce single-stranded DNA on a preparative scale from low amounts of starting templates is necessary for most research involving deoxyribozymes, but is particularly important for performing in vitro selections. While the production of single-stranded RNA is straightforward by means of in vitro transcription, the enzymatic production of single-stranded DNA (ssDNA) on a preparative scale is often difficult. Nevertheless, several methods for the production of ssDNA have been published over the years. Here, we present two PCR methods that we find to be particularly effective, fast, and affordable, which we have adapted for our own needs.
从少量起始模板制备单链 DNA 的能力对于大多数涉及脱氧核酶的研究都是必要的,但对于进行体外选择尤其重要。虽然通过体外转录可以很容易地产生单链 RNA,但在制备规模上酶促产生单链 DNA (ssDNA) 通常是困难的。尽管如此,多年来已经发表了几种制备 ssDNA 的方法。在这里,我们提出了两种我们认为特别有效、快速且经济实惠的 PCR 方法,我们根据自己的需要对其进行了改编。