Efficient microRNA capture and bar-coding via enzymatic oligonucleotide adenylation

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Abstract

Here we report a highly efficient and simplified strategy to preadenylate bar-coded oligonucleotides designed for microRNA (miRNA) capture and multiplex analysis. Using this approach, we enzymatically preadenylated bar-coded oligonucleotides with high efficiency when compared to the chemical method currently used by miRNA investigators. As a case study, we used these oligonucleotides in an ATP-independent ligation to miRNAs, suggesting the utility of our method in end-capture protocols and high-throughput sequencing applications.

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Figure 1: Adenylation of the 3′ adaptor oligonucleotide by T4 DNA ligase.
Figure 2: miRNA capture by ligation.

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Acknowledgements

This work was supported by the Center for Excellence in Genome Sciences grant from the US National Human Genome Research Institute. We thank A.Z. Fire for helpful discussions and assistance in this project. F.V. is supported by a Canadian Institutes of Health Research Fellowship and A.M.S. is supported by the US National Institutes of Health Ruth L. Kirschstein Fellowship.

Author information

Correspondence to Francois Vigneault or George M Church.

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Supplementary Text and Figures

Supplementary Figures 1–2, Supplementary Table 1, Supplementary Methods (PDF 601 kb)

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