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RNA-seq03:21

RNA-seq

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RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases. 
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
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Monitoring Conformational Dynamics of Single Unmodified Proteins using Plasmonic Nanotweezers
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Single Molecule Protein Segments Sequencing by a Plasmonic Nanopore.

Juan Zhou1, Qing Lan1, Wang Li1

  • 1State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210023, China.

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Summary

This study demonstrates how high voltage unfolds single molecule proteins, enabling sequential analysis within a nanopore. This breakthrough enhances protein sequencing capabilities using surface-enhanced Raman scattering (SERS) and nanopore technology.

Keywords:
Protein sequencingSERSplasmonic nanoporesingle molecule detectionunfolding protein

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Area of Science:

  • Biophysics
  • Analytical Chemistry
  • Materials Science

Background:

  • Understanding protein function requires sequential and conformational data.
  • Nanopore electrical detection offers single-molecule protein sensing but struggles with translocation speed and signal quality.
  • Existing methods need improvement for detailed protein analysis.

Purpose of the Study:

  • To develop a method for slowing protein translocation in nanopores.
  • To achieve sequential analysis of single molecule protein segments.
  • To improve signal-to-noise ratio for protein detection in nanopores.

Main Methods:

  • Utilized a surface-enhanced Raman scattering (SERS) active conical gold nanopore.
  • Applied high bias voltage to unfold single molecule (SM) proteins.
  • Recorded sequential SERS traces of different protein segments during translocation.

Main Results:

  • High bias voltage induced protein unfolding, exposing more amino acid residues.
  • Protein unfolding effectively slowed down translocation through the nanopore.
  • Distinct SERS signals were obtained for sequential segments of cytochrome c (cyt c).

Conclusions:

  • Combining single molecule SERS with nanopore technology provides direct insights into protein segments.
  • This approach addresses challenges in nanopore-based protein sequencing.
  • The method shows promise for advancing single molecule protein analysis and sequencing.