Chip-qpcr
Chip-qpcr - Related Videos
10:50
Single-cell Gene Expression Profiling Using FACS and qPCR with Internal Standards
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Cited by 7 •
2017
We describe a method to sort single mammalian cells and to quantify the expression of up to 96 target genes of interest in each cell. This method includes the use of internal qPCR standards to enable the estimation of absolute transcript counts.
09:57
Quantifying Environmental Microorganisms and Viruses Using qPCR
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2023
Source: Laboratories of Dr. Ian Pepper and Dr. Charles Gerba - The University of Arizona Demonstrating Author: Bradley Schmitz Quantitative polymerase chain reaction (qPCR), also known as real-time PCR, is a widely-used molecular technique for enumerating microorganisms in the environment. Prior to this approach, quantifying microorganisms was limited largely to classical culture-based techniques. However, the culturing of microbes from environmental samples can be particularly challenging, and...
09:21
Quantitative Real-Time PCR using the Thermo Scientific Solaris qPCR Assay
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Cited by 3 •
2010
The Solaris qPCR Gene Expression Assays are novel pre-designed qPCR primer/probe combinations designed to simplify the qPCR process without sacrificing the specificity and robustness of the assay.
13:47
Chromatin Immunoprecipitation (ChIP) using Drosophila tissue
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Cited by 10 •
2012
Recently high-throughput sequencing technology has greatly increased sensitivity of Chromatin Immunoprecipitation (ChIP) experiment and prompted its application using purified cells or dissected tissue. Here we delineate a method to use ChIP technique with Drosophila tissue, which can address the endogenous chromatin state in a well-characterized biological system.
12:32
EPA Method 1615. Measurement of Enterovirus and Norovirus Occurrence in Water by Culture and RT-qPCR. Part III. Virus Detection by RT-qPCR
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Cited by 13 •
2016
Here we present a procedure to quantify enterovirus and norovirus in environmental and drinking waters using reverse transcription-quantitative PCR. Mean virus recovery from groundwater with this standardized procedure from EPA Method 1615 was 20% for poliovirus and 30% for murine norovirus.
FAQs
Preserving these interactions maintains the association between a protein or histone modification and the genomic region it occupies. Fragmenting the chromatin afterward creates manageable DNA-protein complexes while retaining those associations for antibody-based recovery. If the interactions are not maintained, the recovered DNA may no longer represent the original regulatory state of the cells.
The antibody determines which chromatin-associated complexes are isolated from the fragmented sample. An antibody directed against a transcription factor focuses the analysis on that factor's DNA associations, whereas one recognizing a histone modification examines a different epigenetic feature. Consequently, antibody choice directly defines the regulatory event evaluated at the selected genomic region.
Quantitative PCR identifies and measures selected DNA sequences within the recovered material. This targeted readout connects the isolated protein or histone modification to defined promoters or regulatory elements rather than evaluating genomic DNA indiscriminately. In cancer studies, the resulting measurements can reveal changes in regulatory associations linked to oncogenes, tumor suppressors, or treatment responses.
A typical workflow preserves protein-DNA interactions, fragments the chromatin, and applies an antibody to isolate complexes containing the target protein or histone modification. DNA is then recovered from the isolated material and analyzed by real-time PCR using selected genomic sequences. These stages link molecular isolation with targeted measurement of regulatory regions.
The method is useful when researchers need to examine transcription-factor binding or epigenetic marks at genomic regions connected with cancer-related gene regulation. Measurements can be compared across normal and tumor cells or between untreated and experimentally treated samples. Such comparisons help assess regulatory mechanisms associated with oncogenes, tumor suppressors, and treatment responses.
Differences in recovered signal at a selected promoter or regulatory element can indicate altered association of the examined protein or histone modification with that region. Comparing cellular conditions therefore supports mechanistic interpretations of gene regulation, including changes related to tumor state or treatment. The approach is especially suited to testing predefined genomic regions of interest.