Sequence Tag Counting

Sequence tag counting is a gene-expression profiling method that measures transcript abundance by sequencing and counting short, identifiable fragments of RNA-derived sequences. In a typical workflow, cellular mRNA is converted to complementary DNA, molecular tags are generated from defined transcript regions, and the tags are sequenced; the number of reads matching each gene provides an abundance estimate. This approach enables quantitative comparison of expression patterns across tissues, developmental stages, treatments, or disease states without requiring prior knowledge of every transcript. It has supported transcriptome analysis, biomarker discovery, and investigations of biological responses at genome scale.

Sequence Tag Counting - Related Videos

Research

JoVE Journal - Biology
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Chromatin Interaction Analysis with Paired-End Tag Sequencing (ChIA-PET) for Mapping Chromatin Interactions and Understanding Transcription Regulation

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Cited by 17 •

2012

Chromatin Interaction Analysis by Paired-End Tag Sequencing (ChIA-PET) is a method for de novo detection of chromatin interactions, for better understanding of transcriptional control.

Education

JoVE Science Education - Basic Biology

Using a Hemacytometer to Count Cells

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2023

Many biomedical experiments require manipulation of a known quantity of cells, in order to achieve accurate, reproducible, and statistically-relevant data. Therefore, learning how to count cells is a particularly essential technique for any successful biomedical scientist. The most common way to count cells is by using a hemacytometer - an instrument that bears two laser-etched grids, which aid in the enumeration of an aliquot cells under a simple light microscope. This data can then be used to...

Counting and Determining the Viability of Cultured Cells

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Cited by 19 •

2008

Determining the number of cells in culture is important in standardization of culture conditions and in performing accurate quantitation experiments. In this video, we demonstrate how cells are counted using a hemacytometer.

Single-cell RNA Sequencing of Fluorescently Labeled Mouse Neurons Using Manual Sorting and Double In Vitro Transcription with Absolute Counts Sequencing (DIVA-Seq)

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Cited by 3 •

2018

This protocol describes the manual sorting procedure to isolate single fluorescently labeled neurons followed by in vitro transcription-based mRNA amplification for high-depth single-cell RNA sequencing.

High-throughput Gene Tagging in Trypanosoma brucei

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Cited by 21 •

2016

Addition of a tag to a protein is a powerful way of gaining insight into its function. Here, we describe a protocol to endogenously tag hundreds of Trypanosoma brucei proteins in parallel such that genome scale tagging is achievable.

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