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DNA sequencing: modular primers for automated walking

L Kotler¹, I Sobolev, L Ulanovsky

¹Department of Structural Biology, Weizmann Institute, Rehovot, Israel.

|September 1, 1994

Summary

This study introduces modular primers for DNA sequencing, significantly reducing time and cost. This advancement enables fully automated DNA sequencing, overcoming previous limitations.

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Area of Science:

Molecular Biology
Genomics
Biotechnology

Background:

Primer synthesis is a bottleneck in DNA sequencing by primer walking.
Current methods require custom primer synthesis for each sequencing walk.
This limits the potential for full automation in DNA sequencing workflows.

Purpose of the Study:

To develop a novel DNA sequencing technology using modular primers.
To eliminate the primer synthesis bottleneck in primer walking.
To enable closed-end automation of the DNA sequencing cycle.

Main Methods:

Modular primers are assembled from short oligonucleotides (5-mers, 6-mers, or 7-mers).
A presynthesized library of approximately 1000 oligonucleotides is utilized.
The modular primers are applied to sequencing with fluorescent dye terminators.

Main Results:

The modular primer approach is expected to reduce sequencing time by 20-50 fold and cost by 5-15 fold.
Achieved success rate and quality of automated sequencing are comparable to conventional primers.
The technique allows for simple purification without protein removal.

Conclusions:

Modular primers offer a significant advancement in DNA sequencing technology.
This approach facilitates cost-effective and time-efficient automated DNA sequencing.
The elimination of custom primer synthesis enables fully automated sequencing cycles.

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