An Efficient Bead‐captured Denaturation Method for Preparing Long 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 4000 nt) 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 20 min, 80 nM of the 200 nt ssDNA strand could be obtained from its duplex DNA. Consider the wide use of single‐stranded DNA substrates of the longer length in the biological and analytical chemistry community, this developed method can circumvent and facilitate the ssDNA preparation process. This can be a practical and handy method for the nucleic acids biochemistry community in preparing long single‐stranded DNA substrates.