The story of oligonucleotide synthesis is a testament to human ingenuity in manipulating the building blocks of life. From single-strand synthesis in the 1960s to today’s high-throughput oligo pools, the journey highlights remarkable technological advancements. These innovations have fueled progress in genomics, synthetic biology, and personalized medicine, making oligonucleotide synthesis indispensable in modern science.

The Birth of Oligonucleotide Synthesis

The history of oligonucleotide synthesis dates back to the 1960s, when scientists first achieved controlled, chemical synthesis of short DNA strands. Early methods relied on the phosphodiester approach, which, though groundbreaking, was limited in efficiency and scalability. The invention of the phosphoramidite method in the 1980s marked a turning point. This approach introduced automated synthesizers, enabling rapid production of DNA sequences with higher precision and yield. For the first time, scientists could design and produce custom oligonucleotides, opening the door to new research possibilities in molecular biology and genetic engineering.

Key Advances in DNA Technology

1. Solid-Phase Synthesis

Solid-phase synthesis, pioneered during the 1980s, revolutionized DNA production by attaching the growing oligonucleotide chain to a solid support. This innovation not only improved the efficiency of synthesis but also simplified purification, making large-scale production feasible.

2. Automated Synthesizers

The automation of DNA synthesis dramatically accelerated the pace of research. Machines could produce oligonucleotides with precise sequences in a fraction of the time required for manual methods, enabling researchers to tackle increasingly complex experiments.

3. High-Fidelity Synthesis

Improvements in reagents and protocols have significantly reduced synthesis errors. Modern phosphoramidite chemistry, combined with enzymatic error correction, ensures that oligonucleotides meet the stringent quality requirements for applications such as gene editing and diagnostics.

The Rise of Complex Oligo Pools

As the demand for high-throughput DNA technologies grew, traditional single-strand synthesis methods faced scalability challenges. Enter oligo pools, which allow for the parallel synthesis of thousands to millions of unique DNA sequences on a single chip. These advancements have been driven by technologies such as:

• Microarray-Based Synthesis: Using photolithography or inkjet printing, microarray platforms can create thousands of oligonucleotides simultaneously. This method has been instrumental in applications like CRISPR library creation and synthetic biology.

• Next-Generation Sequencing: The advent of next-generation sequencing technologies has created a demand for large, diverse oligonucleotide libraries. These libraries are used to generate sequencing libraries, enabling researchers to sequence entire genomes in a single run.

• Synthetic Biology: The field of synthetic biology relies heavily on oligonucleotide synthesis. Complex oligo pools are used to assemble synthetic genes, regulatory elements, and even entire genomes. This has led to a surge in demand for high-quality, custom-synthesized oligonucleotides.

• CRISPR-Cas Gene Editing: CRISPR-Cas technology has revolutionized gene editing, enabling precise and efficient modifications to the genome. Oligo pools play a crucial role in this process, providing the necessary guide RNAs and other components for targeted gene editing.

Continued advancements in DNA technology will drive further innovations in oligo pool synthesis, enabling even more complex and diverse oligo pools. These advancements will have profound implications for a wide range of applications, including personalized medicine, synthetic biology, and biotechnology.

 Applications of Oligo Pool

Synbio Technologies

Oligo Pools Synthesis Services

At Synbio Technologies, our Syno High-Throughput Synthesis Platform utilizes inkjet technology to synthesize up to 890,000 oligonucleotide sequences on a chip in a single parallel run, with a maximum length of 300 nt. During the synthesis process, each reaction site forms a microdroplet isolated from each other, guaranteeing zero cross-contamination, and the synthesized sequences have a high degree of accuracy and homogeneity.

The Customized Oligo Pools Synthesis Service can perfectly match customers' downstream experiments and applications. For example, CRISPR sgRNA screening libraries, high-throughput sequencing, high-throughput gene synthesis, synthetic biology, etc., to improve the efficiency of subsequent high-throughput screening and the success rate of long fragment assembly.

References

Kuiper, B. P., Prins, R. C., & Billerbeck, S. (2022). Oligo pools as an affordable source of synthetic DNA for cost‐effective library construction in protein‐and metabolic pathway engineering. ChemBioChem, 23(7), e202100507.