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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. 
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The first human genome sequencing project cost $2.7 billion and was declared complete in 2003, after 15 years of international cooperation and collaboration between several research teams and funding agencies. Today, with the advent of next-generation sequencing technologies, the cost and time of sequencing a human genome have dropped over 100 fold.
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An Ultrahigh-throughput Microfluidic Platform for Single-cell Genome Sequencing
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Single-cell barcoding and sequencing using droplet microfluidics.

Rapolas Zilionis1,2, Juozas Nainys1, Adrian Veres2,3,4

  • 1Institute of Biotechnology, Vilnius University, Vilnius, Lithuania.

Nature Protocols
|December 9, 2016
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Summary
This summary is machine-generated.

We developed inDrops, a droplet microfluidics method for high-throughput single-cell RNA sequencing. This scalable platform efficiently captures and profiles thousands of cells, revealing cellular heterogeneity in tissues.

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Area of Science:

  • Genomics
  • Molecular Biology
  • Biotechnology

Background:

  • Single-cell RNA sequencing (scRNA-seq) is crucial for understanding cellular heterogeneity.
  • Existing high-throughput methods are needed for unbiased cell diversity capture.
  • Droplet microfluidics offers a promising avenue for massively parallel single-cell analysis.

Purpose of the Study:

  • To detail the inDrops method for high-throughput single-cell RNA sequencing.
  • To provide a robust and scalable platform for capturing and profiling large numbers of cells.
  • To enable comprehensive analysis of cellular heterogeneity in various biological samples.

Main Methods:

  • Encapsulation of cells with barcoding hydrogel beads in nanoliter droplets.
  • Single-cell lysis and barcoding of mRNA via reverse transcription (RT).
  • RNA-seq library preparation for downstream analysis.

Main Results:

  • The inDrops platform can index over 15,000 cells per hour.
  • It achieves high cell capture efficiency (>75%), even in small samples.
  • The method is robust, scalable, and minimizes reagent use.

Conclusions:

  • inDrops is a powerful tool for high-throughput single-cell transcriptomics.
  • It enables efficient profiling of thousands to tens of thousands of cells.
  • This platform significantly advances the study of cellular heterogeneity in health and disease.