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Global Identification of Co-Translational Interaction Networks by Selective Ribosome Profiling
Published on: October 7, 2021
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A rapid protocol for ribosome profiling of low input samples
Andreas Meindl1, Markus Romberger1, Gerhard Lehmann2
1Regensburg Center for Biochemistry, University of Regensburg, Regensburg, Germany.
Nucleic Acids Research
|May 29, 2023
Summary
We developed an ultra-rapid ribosome profiling method for low-input samples. This new protocol enables high-quality translation analysis from small amounts of biological material.
Area of Science:
- Molecular Biology
- Genomics
- Translational Control
Background:
- Ribosome profiling offers high-resolution insights into cellular translation.
- Traditional ribosome profiling is complex, time-consuming, and requires large sample inputs, limiting its application.
Purpose of the Study:
- To develop an ultra-rapid ribosome profiling protocol.
- To enable high-quality translation analysis from low-input samples.
Main Methods:
- Developed a streamlined sequencing library preparation protocol.
- Utilized solid-phase purification of reaction intermediates.
- Reduced RNA input requirement to as little as 0.1 pmol.
Main Results:
- Achieved a one-day turnaround for sequencing library preparation.
- Demonstrated successful ribosome profiling with significantly reduced sample input.
- Maintained high sensitivity and data quality.
Conclusions:
- The new protocol significantly simplifies and accelerates ribosome profiling.
- Enables ribosome profiling analysis on small or limited samples.
- Expands the applicability and accessibility of ribosome profiling techniques.
