Duplex Ish
Duplex Ish - Related Videos
11:42
Iterative Optimization of DNA Duplexes for Crystallization of SeqA-DNA Complexes
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2012
Crystal structure of protein–DNA complexes can provide insight into protein function, mechanism, as well as, the nature of the specific interaction. Here, we report how to optimize the length, sequence and ends of duplex DNA for co-crystallization with Escherichia coli SeqA, a negative regulator of replication initiation.
04:10
Helicase Activity Measurement of a Target Protein Using Biotin-Labeled RNA Duplexes
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2025
In this video, we demonstrate the procedure to determine the helicase activity of a target protein to unwind the biotin-labeled dsRNA substrate. The activity of the enzyme was identified by analyzing the electrophoretic mobility shift, followed by a chemiluminescence assay using chemiluminescent enzyme-conjugated streptavidin.
08:45
Duplex Digital PCR for Simultaneous Quantification of Dual Genetic Markers
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2025
In this video, we demonstrate the duplex digital PCR (ddPCR) technique — a modification of the traditional PCR technique that is useful in detecting two different genetic markers simultaneously. A single PCR reaction is partitioned into nanoliter-sized emulsified droplets that are independently amplified, and the detection of differently colored fluorescence amplification signals from the fraction of droplets is used to compute the initial concentration of the target sequences.
12:14
A Duplex Digital PCR Assay for Simultaneous Quantification of the Enterococcus spp. and the Human Fecal-associated HF183 Marker in Waters
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Cited by 14 •
2016
This manuscript describes a duplex digital PCR assay that can be used to simultaneously quantify Enterococcus spp. and the HF183 genetic markers as indicators of general and human-associated fecal contamination in recreational waters.
10:41
Analysis of Cerebral Vasospasm in a Murine Model of Subarachnoid Hemorrhage with High Frequency Transcranial Duplex Ultrasound
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2021
The aim of this manuscript is to present a sonography-based method that allows in vivo imaging of blood flow in cerebral arteries in mice. We demonstrate its application to determine changes in blood flow velocities associated with vasospasm in murine models of subarachnoid hemorrhage (SAH).
FAQs
Each transcript is queried with a sequence-specific probe linked to a different detectable label. Because the labels generate separate signals, investigators can assign each spatial pattern to its corresponding RNA target rather than treating the specimen as a single combined measurement. This distinction is essential for evaluating whether transcripts occupy the same cells or different locations.
Co-expression identifies cells or tissue locations containing both RNA transcripts in the same specimen. That combined pattern can reveal molecularly defined populations that would be difficult to distinguish from either marker independently. In neuroscience, the result helps relate gene-expression combinations to neuronal or glial identity and provides more specific information about cellular organization.
Spatial context connects transcript detection to the cells and brain regions where signals occur. This allows researchers to compare regional expression patterns and determine whether two markers overlap within a population or occupy separate populations. Preserving that relationship reduces ambiguity that can arise when transcript measurements are analyzed without their original tissue location.
The analysis begins with intact cells or tissue sections and sequence-specific probes selected for the two RNA targets. Each probe hybridizes to its complementary transcript, after which the separate detectable labels are used to visualize the resulting signals. Interpreting the specimen then focuses on signal location, overlap, and distribution across cells or tissue regions.
By examining two transcript patterns together, investigators can locate molecular markers across brain regions and identify neuronal or glial populations with particular expression combinations. The tissue-based readout preserves regional organization, so molecular classifications can be considered alongside anatomical distribution. This supports analysis of how cellular identities are arranged within neural tissue.
Duplex ISH can contribute to studies of circuit organization, cell identity, and disease-related gene-expression changes. Researchers may use paired transcript localization to examine how molecularly defined cells are distributed within brain regions or associated with circuit structure. Comparing these patterns across specimens can also help identify changes linked to neurological disease states.