Enzymatic synthesis of single-stranded DNA by in vitro transcription

Efficient ways to produce single-stranded DNA are of great interest for diverse applications in molecular biology and nanotechnology. In the present study, we select T7 RNA polymerase mutants with reduced substrate specificity to employ in vitro transcription reaction for the synthesis of DNA oligonucleotides, either individually or in pools. Transcribed oligonucleotides can serve as PCR primers which makes the present approach a platform for high-quality, simple, and cheap oligonucleotide synthesis.