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Related Concept Videos

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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An Ultrahigh-throughput Microfluidic Platform for Single-cell Genome Sequencing
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Single-cell isolation by a modular single-cell pipette for RNA-sequencing.

Kai Zhang1, Min Gao2, Zechen Chong3

  • 1Department of Nanomedicine, Houston Methodist Research Institute, Houston, TX 77030, USA. lqin@houstonmethodist.org and Department of Cell and Developmental Biology, Weill Medical College of Cornell University, New York, NY 10065, USA.

Lab on a Chip
|November 15, 2016
PubMed
Summary

Researchers developed a modular single-cell pipette (mSCP) for efficient live single-cell isolation. This tool enables rapid, high-efficiency cell capture for downstream transcriptome analysis.

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Genomics

Background:

  • Single-cell transcriptome sequencing demands efficient methods for isolating live cells into containers like PCR tubes.
  • Current methods may lack convenience, speed, or reliability for high-throughput applications.

Purpose of the Study:

  • To develop and validate a novel, modular single-cell pipette (mSCP) for rapid and efficient isolation of live single cells.
  • To assess the mSCP's compatibility with microscopic identification and its suitability for downstream molecular analyses.

Main Methods:

  • The modular single-cell pipette (mSCP) was designed with three components: SCP-Tip, air-displacement pipette (ADP), and ADP-Tips.
  • The SCP-Tip incorporates a hydrodynamic trap for cell capture.
  • The mSCP allows for easy assembly, disassembly, and microscopic identification of captured cells.

Main Results:

  • The mSCP successfully isolates single cells from suspensions at concentrations of 5-10 cells/μL.
  • Microscopic identification ensures 100% single-cell isolation efficiency.
  • Isolated live single cells are contained in submicroliter volumes, suitable for PCR and RNA sequencing.

Conclusions:

  • The modular single-cell pipette (mSCP) offers a convenient, rapid, and highly efficient solution for live single-cell isolation.
  • The mSCP is a powerful tool for preparing samples for single-cell transcriptome analysis.