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A Rapid, Cost-Effective Method of Assembly and Purification of Synthetic DNA Probes >100 bp

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  • Michael A Jensen
  • Lauren Jauregui
  • Ronald W Davis

Abstract

Here we introduce a rapid, cost-effective method of generating molecular DNA probes in just under 15 minutes without the need for expensive, time-consuming gel-extraction steps. As an example, we enzymatically concatenated six variable strands (50 bp) with a common strand sequence (51 bp) in a single pool using Fast-Link DNA ligase to produce 101 bp targets (10 min). Unincorporated species were then filtered out by passing the crude reaction through a size-exclusion column ( 12) could be achieved with further optimization. Moreover, for large-scale assays, we envision this method to be fully automated with the use of robotics such as the Biomek FX; here, potentially thousands of samples could be pooled, ligated and purified in either a 96, 384 or 1536-well platform in just minutes.

Suggested Citation

  • Michael A Jensen & Lauren Jauregui & Ronald W Davis, 2012. "A Rapid, Cost-Effective Method of Assembly and Purification of Synthetic DNA Probes >100 bp," PLOS ONE, Public Library of Science, vol. 7(4), pages 1-4, April.
    • Handle: RePEc:plo:pone00:0034373
    DOI: 10.1371/journal.pone.0034373
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