Transcript Detection
Transcript Detection - Related Videos
04:31
Electrophoretic Mobility Shift Assay for Detection of Transcription Factor-DNA Complexes
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2025
In this video, we demonstrate the electrophoretic mobility shift assay to detect protein-DNA interactions using infrared fluorescent-labeled DNA probes. When the protein binds to the DNA probe, it forms a large complex that migrates slowly on and appears as a shifted band on the gel.
07:50
Chip-Based Digital PCR to Detect Rare Transcript Variants Using a Nanofluidic Chip
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2025
This video demonstrates chip-based digital PCR — a variation of the digital PCR technique that is useful in detecting rare transcript variants. The PCR reaction is partitioned into the chambers of a nanofluidic chip, each of which acts as an independent reaction. The detection of fluorescence signals from the chambers with amplified targets confirms the presence of rare transcript variants in the sample.
12:12
Analysis of Termination of Transcription Using BrUTP-strand-specific Transcription Run-on (TRO) Approach
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Cited by 10 •
2017
We describe a basic experimental approach for analysis of termination of transcription by RNA polymerase II in vivo using BrUTP by the strand-specific transcription run-on (TRO) approach in budding yeast. This protocol can be extended to study transcription termination by other RNA polymerases both in yeast and higher eukaryotes.
02:16
Transcription Factors
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2019
Tissue-specific transcription factors contribute to diverse cellular functions in mammals. For example, the gene for beta globin, a major component of hemoglobin, is present in all cells of the body. However, it is only expressed in red blood cells because the transcription factors that can bind to the promoter sequences of the beta globin gene are only expressed in these cells. Tissue-specific transcription factors also ensure that mutations in these factors may impair only the function of...
03:45
Antibody Staining of Scale Melanocytes: A Technique to Detect Transcription Factors Expressed in Dorsal Scale-associated Melanocytes of Zebrafish Models
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2023
This video describes the antibody staining method to detect transcription factors expression in dorsal scale-associated melanocytes of zebrafish models. This immunostaining technique helps specifically identify the melanocyte-inducing transfection factor-A in the transgenic zebrafish.
FAQs
Sequence complementarity allows a probe, primer, or sequencing adapter to recognize a matching nucleotide sequence within the RNA-derived target. Nonmatching sequences are less likely to support the intended recognition step, helping associate the resulting signal with a particular transcript. This sequence-based selectivity enables researchers to distinguish gene-expression patterns among cells, tissues, developmental stages, or experimental conditions.
These components support different stages of the measurement process. Labeled probes recognize target transcripts through hybridization, primers support amplification of selected sequences, and sequencing adapters enable target molecules to be processed for nucleotide-sequence readout. Choosing among them determines whether the experiment emphasizes direct recognition, signal generation through amplification, or sequence-based analysis of RNA molecules.
Hybridization detects a target through complementary pairing with a labeled probe, whereas amplification increases the detectable representation of selected sequences before measurement. Sequencing-based readout instead provides information from nucleotide sequences associated with the transcripts. These approaches can therefore emphasize target recognition, measurable signal production, or sequence-level characterization, depending on the biological question.
Transcript abundance provides an indication of how much of a particular RNA molecule is present under a given biological condition. Comparing abundance among developmental stages, treatments, or genetic and environmental perturbations can reveal changes in gene activity. Such comparisons help connect altered RNA levels with cellular regulation, disease-associated states, or responses to treatment.
A typical workflow begins by selecting the transcript or sequence of interest, followed by applying a complementary probe, primer, or sequencing adapter. The chosen recognition strategy then produces a detectable signal through hybridization, amplification, or nucleotide-sequence readout. Measurements are compared across the relevant cells, tissues, developmental stages, or experimental conditions to identify expression differences.
Researchers would use this approach when they need evidence that gene activity differs between defined biological states. Examples supported by the method include comparisons across developmental stages, disease and treatment conditions, or cells exposed to environmental or genetic perturbations. The resulting measurements can show whether selected transcripts increase, decrease, or vary between those contexts.
Because recognition and readout depend on nucleotide sequences, transcript detection can help distinguish related RNA forms when their sequences differ in relevant regions. Measuring these forms separately can extend expression analysis beyond total transcript abundance and support investigations of RNA diversity. In biology, this information contributes to understanding transcript regulation and changes associated with cellular or disease states.