An Efficient Bead-captured Denaturation Method for Preparing Long Single-stranded DNA

作者：Kao Wei Siang; Li Hung Wen^*

来源：Journal of the Chinese Chemical Society, 2017, 64(9): 1065-1070.

DOI：10.1002/jccs.201700142

• Alkaline denaturation
• Bead capturing
• Single-stranded DNA

Previous methods to prepare single-stranded DNA (ssDNA) substrates are limited to short DNA lengths and inefficient. We have developed an efficient and rapid method to prepare long ssDNA substrates (up to 4000nt) based on the denaturation of the bead-captured DNA substrates, with the individual steps optimized. Immobilization of the targeted DNA substrates on the antibody-modified beads allows easy separation of the denatured targeted ssDNA strand. This method also allows the recovery of the captured strand, making it possible to obtain two ssDNA strands from the same duplex DNA. Within 20min, 80nM of the 200nt ssDNA strand could be obtained from its duplex DNA.

• 出版日期2017-9