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Small RNA Library Preparation Method for Next-Generation Sequencing Using Chemical Modifications to Prevent Adapter
Sabrina Shore1, Jordana M Henderson1, Alexandre Lebedev1
1Research and Development, Cell and Molecular Biology, TriLink BioTechnologies LLC., San Diego, California, United States of America.
Plos One
|November 23, 2016
Summary
This study introduces a novel small RNA (sRNA) library preparation method using modified adapters to reduce adapter dimer formation. This approach lowers sample input needs and removes the need for gel purification, enabling automated sRNA sequencing.
Area of Science:
- Molecular Biology
- Genomics
- Biotechnology
Background:
- Automation of RNA and DNA sample preparation facilitates high-throughput next-generation sequencing (NGS).
- Small RNA (sRNA) sequencing adoption is limited by high sample input requirements and adapter dimer formation during library preparation.
- Adapter dimers are size-similar to tagged libraries, necessitating gel purification, which is inefficient at low sample inputs.
Purpose of the Study:
- To develop a more specific sRNA library preparation workflow.
- To overcome limitations of high sample input and adapter dimer contamination in sRNA sequencing.
- To enable lower sample input and eliminate gel purification for automated sRNA library preparation.
Main Methods:
- Development of a novel library preparation approach utilizing modified adapters.
- The modified adapters are designed to suppress the formation of adapter dimers.
- Evaluation of the workflow for its ability to reduce sample input and eliminate gel purification.
Main Results:
- The novel workflow significantly suppresses adapter dimer formation.
- This suppression allows for reduced sample input requirements.
- The elimination of the gel purification step was achieved.
Conclusions:
- The modified adapter approach enhances the specificity of sRNA library preparation.
- This method addresses key limitations hindering broader sRNA sequencing adoption.
- The workflow is automatable, paving the way for high-throughput, sensitive sRNA analysis.
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