Researchers at the Weizmann Institute developed a novel method to design error-free DNA libraries from error-prone oligonucleotides. The system surpasses existing methods for de novo synthesis of DNA libraries in speed, precision, amenability to automation and ease of combining synthetic with natural DNA fragments.
All DNA construction protocols struggle with the cumbersome task of cloning and sequencing synthetic DNA fragments, seeking an error-free one. The problem is worsened for longer synthetic DNA which is more prone to errors. Time spent on error correction, clone selection and sequencing is a major bottleneck that prevents de novo DNA synthesis from becoming a routine procedure in labs.
This innovative solution significantly decreases the need for labor-intensive time-consuming error correction methods, cloning and sequencing. Furthermore, efficient editing and reassembly of different genes is made possible due to a smart recursive reconstruction process.
- Design and construction of synthetic biological molecules and organisms.
- Construction of designer DNA libraries.
- Applicable in any lab with standard lab equipment. Faster and more precise than existing methods.
- Amenable to automation, full synthesis in vitro with a modified smPCR protocol.
- Very simple to combine synthetic and natural DNA fragments.
- Does not require additional or external methods or reagents for error correction
Divide and Conquer (D&C), the quintessential recursive problem-solving technique, is applied in silico to divide the target DNA sequence into overlapping oligonucleotides short enough to be synthesized directly, albeit with errors; error-prone oligonucleotides are recursively combined in vitro, forming error-prone DNA molecules; error-free fragments of these molecules are then identified, extracted and used as new, typically longer and more accurate, inputs to another iteration of the recursive construction procedure; the entire process repeats until an error-free target molecule is formed.