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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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RNA Stability01:53

RNA Stability

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Intact DNA strands can be found in fossils, while scientists sometimes struggle to keep RNA intact under laboratory conditions. The structural variations between RNA and DNA underlie the differences in their stability and longevity. Because DNA is double-stranded, it is inherently more stable. The single-stranded structure of RNA is less stable but also more flexible and can form weak internal bonds. Additionally, most RNAs in the cell are relatively short, while DNA can be up to 250 million...
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RNA Interference01:23

RNA Interference

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RNA interference (RNAi) is a process in which a small non-coding RNA molecule blocks the post-transcriptional expression of a gene by binding to its messenger RNA (mRNA) and preventing the protein from being translated.
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
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Alternative RNA Splicing02:18

Alternative RNA Splicing

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Alternative RNA splicing is the regulated splicing of exons and introns to produce different mature mRNAs from a single pre-mRNA. Unlike in constitutive splicing where a single gene produces a single type of mRNA, alternative splicing allows an organism to produce multiple proteins from a single gene and plays an important role in protein diversity.
There are five types of alternative RNA splicing that vary in the ways the pre-mRNA segments are removed or retained in the mature mRNA. The first...
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RNA Splicing01:32

RNA Splicing

60.5K
Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...
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Types of RNA01:23

Types of RNA

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Overview
Three main types of RNA are involved in protein synthesis: messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). These RNAs perform diverse functions and can be broadly classified as protein-coding or non-coding RNA. Non-coding RNAs play important roles in the regulation of gene expression in response to developmental and environmental changes. Non-coding RNAs in prokaryotes can be manipulated to develop more effective antibacterial drugs for human or animal use.
RNA...
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Identification of Footprints of RNA:Protein Complexes via RNA Immunoprecipitation in Tandem Followed by Sequencing RIPiT-Seq
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Identification of Footprints of RNA:Protein Complexes via RNA Immunoprecipitation in Tandem Followed by Sequencing RIPiT-Seq

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Single-Cell RNA Sequencing with Drop-Seq.

Josephine Bageritz1, Gianmarco Raddi2,3,4

  • 1Division Signaling and Functional Genomics, German Cancer Research Center (DKFZ), Heidelberg, Germany. j.bageritz@dkfz-heidelberg.de.

Methods in Molecular Biology (Clifton, N.J.)
|April 28, 2019
PubMed
Summary
This summary is machine-generated.

Drop-Seq is a cost-effective, high-throughput method for single-cell transcriptome profiling. This technique encapsulates cells and barcoded beads in droplets for efficient mRNA capture and sequencing.

Keywords:
Cell barcodingDrop-seqDroplet technologyMicrofluidicsSingle-cell RNA-sequencingSystems biologyTranscriptomics

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Single-cell RNA-Seq of Defined Subsets of Retinal Ganglion Cells
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Gel-seq: A Method for Simultaneous Sequencing Library Preparation of DNA and RNA Using Hydrogel Matrices
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Single-cell RNA-Seq of Defined Subsets of Retinal Ganglion Cells
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Single-cell RNA-Seq of Defined Subsets of Retinal Ganglion Cells

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Area of Science:

  • Molecular Biology
  • Genomics
  • Biotechnology

Background:

  • Single-cell analysis is crucial for understanding cellular heterogeneity.
  • Existing methods can be costly and low-throughput.
  • There is a need for accessible platforms for large-scale single-cell transcriptomics.

Purpose of the Study:

  • To introduce and describe the Drop-Seq platform.
  • To demonstrate its capability for high-throughput single-cell RNA sequencing.
  • To provide a cost-effective solution for profiling thousands of cells.

Main Methods:

  • Cells and barcoded microparticles are co-encapsulated in droplets using microfluidics.
  • Cell lysis and RNA hybridization occur within individual droplets.
  • Post-droplet processing includes reverse transcription, PCR, and sequencing of barcoded mRNA.

Main Results:

  • Enables simultaneous profiling of thousands of single-cell transcriptomes.
  • Maintains cellular origin information for each transcript.
  • Provides a scalable and cost-effective approach to single-cell analysis.

Conclusions:

  • Drop-Seq is a powerful platform for high-throughput single-cell RNA sequencing.
  • It significantly reduces the cost and complexity of single-cell transcriptomic studies.
  • Facilitates deeper insights into cellular heterogeneity and biological systems.