Barcode-Guided Nucleotide Capture for High-Purity DNA Synthesis
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Solution Overview
Problem
Conventional DNA synthesis methods result in high error rates and costly manual separation processes, limiting the efficiency and cost-effectiveness of DNA sequence synthesis and purification.
Innovation Solution
A method involving preparation of nucleic acid fragments, tagging with barcode sequences, determining location coordinates, and retrieving validated fragments using a support-based apparatus with a parallel sequencer, imager, and extractor to achieve high-purity nucleic acid capture.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If chemical synthesis methods are used for DNA sequence synthesis, then cost is reduced, but error rate increases significantly
Solution Approach 1:
The DNA synthesis process is divided into two independent stages: (1) parallel chemical synthesis of many DNA fragments simultaneously on a microarray, and (2) selective amplification and purification of only the error-free fragments through barcode tagging and PCR. This segmentation allows cheap chemical synthesis to coexist with high accuracy selection.
Solution Approach 2:
The method incorporates feedback through sequencing validation of barcode sequences to identify and select only error-free DNA fragments. The sequencing results feed back into the selection process, allowing iterative purification until high purity is achieved, thus resolving the accuracy-cost contradiction.
2Ease of manufacture
If manual separation methods are used to purify DNA sequences, then cost is reduced, but time consumption increases significantly
Solution Approach 1:
Barcode sequences are attached to DNA fragments during the synthesis process itself, before separation is needed. This preliminary tagging enables automated identification and selection later, eliminating the need for time-consuming manual separation while maintaining low cost through automation of the selection process.
Solution Approach 2:
The manual mechanical process of separating DNA fragments is replaced with an automated system using barcode sequencing and computational selection. The mechanical separation is substituted by information-based identification through barcode reading and automated PCR amplification of selected fragments, dramatically reducing time while keeping costs low.
3Ease of manufacture
If high-throughput sequencing is applied to microchip oligonucleotides, then sequencing cost is reduced, but difficulty of isolating target DNA increases
Solution Approach 1:
Barcode sequences serve as an intermediary element that links the synthesized DNA fragments to their sequence information. The barcodes enable high-throughput sequencing of individual fragments while simultaneously providing a handle for their automated identification and isolation, thus resolving the contradiction between low sequencing cost and difficult target isolation.
Data Source
AI summary
The present invention relates to a technique for synthesizing DNA sequences in high purity at low cost. More specifically, the present invention relates to a method and apparatus for capturing high-purity nucleotides that is suitable for use in the field of genome/gene synthesis.