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Updated: Dec 22, 2025

A Droplet-Based Microfluidic Approach and Microsphere-PCR Amplification for Single-Stranded DNA Amplicons
Published on: November 14, 2018
Construction of a system for single-stranded DNA isolation
Min Hao1,2,3, Huanbang Huang1,2,3, Yasai Hu1,2,3
1School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, China.
Objective:
The system of Strep-Tactin and StrepII tag-SSB proteins binding (ST-SSB) was established to isolate the purified single-stranded DNA in a single step with low cost and high efficiency.
Results:
We demonstrate that in the presence of large amounts of dsDNA, the ssDNA binding specificity of Escherichia coli (E. coli) single stranded DNA binding (EcSSB) protein was stronger than gene-5-protein (g5p). ST-SSB system relies on the affinity between Strep-Tactin, StrepII tag-SSB protein and ssDNA in binding buffer. Here, we successfully isolated the purified ssDNA from mixed DNA (ds- and ss-DNA form) samples and asymmetric polymerase chain reaction (aPCR) products. This system can purify ssDNA in a single tube within 1 h, and the recovery efficiency of purified ssDNA was around 60%.
Conclusions:
The ST-SSB system has obvious advantages of high efficiency and one-step purification to recycle any ssDNA.
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