Immobilized MutS-Mediated Error Removal of Microchip-Synthesized DNA

Abstract

Applications of microchip-synthesized oligonucleotides for de novo gene synthesis are limited primarily by their high error rates. The mismatch binding protein MutS, which can specifically recognize and bind to mismatches, is one of the cheapest tools for error correction of synthetic DNA. Here, we describe a protocol for removing errors in microchip-synthesized oligonucleotides and for the assembly of DNA segments using these oligonucleotides. This protocol can also be used in traditional de novo gene DNA synthesis.

Current and Emerging Methods for the Synthesis of Single-Stranded DNA

“…The product yields of array-based oligo synthesis are typically at the femtomolar scale, i.e., two to four orders of magnitude lower than traditional column-based synthesis [29]. There have been many attempts to increase the scale, quality, and quantity of multi-channel synthesized oligonucleotides [50,51]. Continued improvements in array design, along with the optimization of synthesis reagents and processes, will hopefully deliver platforms for the synthesis of high-quality oligo sequences from arrays, potentially establishing them as the go-to source for multi-channel oligo sequence for gene synthesis applications.…”