Quantitative Pcr
Quantitative Pcr - Related Videos
09:26
Cerebrospinal Fluid MicroRNA Profiling Using Quantitative Real Time PCR
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Cited by 10 •
2014
We describe a protocol of real time PCR to profile microRNAs in the cerebrospinal fluid (CSF). With the exception of RNA extraction protocols, the procedure can be extended to RNA extracted from other body fluids, cultured cells, or tissue specimens.
08:30
MicroRNA Detection in Prostate Tumors by Quantitative Real-time PCR (qPCR)
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Cited by 18 •
2012
Quantitative Real Time polymerase chain reaction (qPCR) is a rapid and sensitive method to investigate the expression levels of various microRNA (miRNA) molecules in tumor samples. Using this method expression of hundreds of different miRNA molecules can be amplified, quantified, and analyzed from the same cDNA template.
09:03
Profiling Individual Human Embryonic Stem Cells by Quantitative RT-PCR
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Cited by 4 •
2014
Single cell gene expression assay is needed for understanding stem cell heterogeneities.
10:28
Probe-based Real-time PCR Approaches for Quantitative Measurement of microRNAs
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Cited by 33 •
2015
Circulating microRNAs have recently emerged as promising and novel biomarkers for various cancers and other diseases. The goal of this article is to discuss three different probe-based real-time PCR platforms and methods that are available to quantify and determine the abundance of circulating microRNAs.
04:08
Real-Time Quantitative Reverse Transcription PCR for Diagnosing Viral Infections
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2025
This video demonstrates a quantitative method for detecting viral infection using real-time reverse transcription PCR. The reverse transcription step converts the RNA to cDNA, which is then amplified via PCR cycles. The presence of viral RNA in the sample is confirmed by analyzing the amplification and dissociation curves of DNA obtained from the PCR.
FAQs
The Ct value marks the cycle at which fluorescence becomes detectable and connects that point with the amount of template present at the start. A sample reaching the detection threshold earlier generally contains more starting target than one requiring additional cycles. Comparing Ct values therefore supports relative estimates of genetic material across biological samples.
Both approaches generate fluorescence as amplified DNA accumulates, but they report that accumulation differently. A sequence-specific probe produces signal in relation to the target sequence, whereas an intercalating dye responds to amplified DNA. This distinction matters when selecting how specifically the assay should monitor the sequence of interest.
The three stages support successive rounds of target amplification. Denaturation separates DNA strands, primer annealing allows primers to bind their complementary targets, and extension enables new DNA synthesis. Repeating these stages increases the amount of the selected sequence while fluorescence tracks product accumulation, creating a quantitative signal during the reaction.
qPCR provides a way to measure selected genetic targets while examining biological samples, making it useful for gene expression analysis. Researchers can compare quantitative signals among samples to investigate cellular responses or changes associated with experimental conditions. Its speed and sensitivity also support analysis when many samples or specific targets must be evaluated.
The method is useful when researchers need to determine whether a specific genetic sequence associated with a pathogen is present and to estimate the amount of target material. The same quantitative capability supports monitoring biological samples over time or across conditions, helping investigators track changes in detected genetic material rather than relying only on presence or absence.
For genotyping, qPCR can examine selected genetic sequences to help distinguish or assess genetic states in biological samples. It can also validate sequencing or other molecular assay results by providing an independent quantitative measurement of a specified target. These applications extend qPCR beyond expression studies and connect targeted measurements with broader molecular analyses.