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Related Experiment Video

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Multiplexed Single Cell mRNA Sequencing Analysis of Mouse Embryonic Cells
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Multiplexed optical coding nanobeads and their application in single-molecule counting analysis for multiple gene

Lu Li1, Liping Shen1, Xiaoqian Zhang2

  • 1School of Chemistry and Chemical Engineering, Shandong University, Jinan 250100, China.

Analytica Chimica Acta
|September 1, 2015
PubMed
Summary
This summary is machine-generated.

Researchers developed novel multiplexed optical coding nanobeads (MOCNBs) for precise gene expression analysis. These nanobeads enable accurate single-molecule counting and multi-gene expression analysis in cells.

Keywords:
Deoxyribonucleic acid determinationMultiple gene expressionMultiplexed optical coding nanobeadsSingle-molecule counting analysis

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Area of Science:

  • Nanotechnology
  • Molecular Biology
  • Biophysics

Background:

  • Accurate quantification of gene expression is crucial for understanding cellular processes.
  • Existing methods for multi-gene analysis can be complex and lack single-molecule resolution.

Purpose of the Study:

  • To develop a novel method for fabricating multiplexed optical coding nanobeads (MOCNBs).
  • To demonstrate the application of MOCNBs in single-molecule counting analysis (SMCA) for multi-gene expression analysis (MGEA).

Main Methods:

  • Fabrication of MOCNBs by hybridizing coding DNAs with different fluorescent dyes (Cy5, FAM, AMCA) to magnetic nanobeads.
  • Utilizing overlapping fluorescence images for MOCNB identification and color coding.
  • Applying SMCA-based MGEA to quantify multiple messenger RNAs (mRNAs) via complementary DNAs (cDNAs).

Main Results:

  • Successfully developed MOCNBs with distinct colors observable under fluorescence microscopy.
  • Demonstrated simultaneous quantification of three gene expression profiles in Helicoverpa armigera cells.
  • Validated the SMCA-based MGEA technique for analyzing gene expression changes in response to steroid hormone induction.

Conclusions:

  • MOCNBs offer a robust platform for multiplexed optical coding.
  • The SMCA-based MGEA technique provides a sensitive method for single-molecule gene expression quantification.
  • This approach has potential applications in various fields of molecular biology and diagnostics.