Microdroplet Pcr
Microdroplet Pcr - Related Videos
08:20
Capillary-based Centrifugal Microfluidic Device for Size-controllable Formation of Monodisperse Microdroplets
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Cited by 9 •
2016
Here, we demonstrate a simple production method for size-controllable, monodisperse, water-in-oil (W/O) microdroplets using a capillary-based centrifugal microfluidic device. This method requires only a small sample volume and enables high-yield production. We expect this method will be useful for rapid biochemical and cellular analyses.
03:20
Real-Time Monitoring of Bacterial Growth Using a Microfluidic Microdroplet Culture System
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2025
Source: Jian, X. et. al., Automated Microbial Cultivation and Adaptive Evolution using Microbial Microdroplet Culture System (MMC). J. Vis. Exp. (2022)This video demonstrates the use of a microfluidic droplet-based platform for real-time monitoring of bacterial growth. By generating uniform droplets containing bacterial suspensions and guiding them through a detection pathway, the system enables continuous optical measurement of cell density to construct a growth curve.
03:42
Adaptive Evolution of Bacteria Using a Microbial Microdroplet Culture System
0 Views •
2025
Source: Jian, X., et al., Automated Microbial Cultivation and Adaptive Evolution using Microbial Microdroplet Culture System (MMC). J. Vis. Exp. (2022).This video demonstrates the use of a microbial microdroplet culture system to drive adaptive evolution of bacteria toward methanol tolerance. By mixing engineered bacterial cultures with a high-methanol stress medium and segmenting them into microdroplets, the system selects for mutants that exhibit improved growth under methanol stress. Through...
08:51
Genotyping of Staphylococcus aureus by Ribosomal Spacer PCR (RS-PCR)
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Cited by 9 •
2016
Here, ribosomal spacer PCR (RS-PCR) is used together with a miniaturized electrophoresis system as a fast and high resolution method for genotyping S. aureus at moderate costs allowing a high throughput.
03:41
Agarose Gel Electrophoresis of DNA Amplicons Post PCR: A Method to Analyze Products of Multiplex PCR and Evaluate PCR Reaction Success
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2025
In this video, we demonstrate the separation of bacterial PCR-amplified DNA using agarose gel electrophoresis. Agarose gel functions as a molecular sieve, enabling the negatively-charged DNA to migrate based upon their size under an applied electric field.
FAQs
The quantitative result comes from counting fluorescence-positive and fluorescence-negative droplets after amplification. Positive droplets indicate that the target sequence was amplified, whereas negative droplets provide the complementary measurement needed for statistical estimation. Applying a Poisson distribution to these counts converts the observed pattern into an estimate of the original target copy number, rather than a simple signal intensity.
Partitioning makes concentration differences easier to resolve because the sample is assessed across many separate reaction chambers. This supports detection when a target is present at low abundance or mixed with complex material. In cancer studies, that sensitivity is particularly useful for identifying rare mutation signals, copy-number changes, or circulating tumor DNA within the broader nucleic-acid sample.
Fluorescence supplies the classification used for downstream quantification. After thermal cycling, each droplet is assigned as containing an amplified target or not, and the balance between these categories determines the statistical estimate. Consequently, interpretation depends on analyzing the complete positive-and-negative droplet pattern rather than considering only droplets that display a fluorescence signal.
Cancer samples may contain targets at very different concentrations, making small differences relevant to biomarker analysis. Microdroplet PCR addresses this need by tolerating small concentration differences and producing a quantitative copy-number estimate. That combination supports measurements involving rare mutations, circulating tumor DNA, and molecular changes followed during treatment.
An assay proceeds from partitioning the DNA sample into droplets to thermal cycling, followed by fluorescence-based droplet assessment. The post-cycling readout separates droplets with amplified target sequences from those without them. Statistical analysis then uses both groups to estimate starting copy number, linking the reaction steps to a quantitative nucleic-acid result.
In cancer research, the resulting measurements can address whether a rare mutation is detectable, whether a copy-number change is present, and whether circulating tumor DNA can be measured in a complex sample. The same analytical approach also supports treatment monitoring and minimal residual disease assessment, where sensitive biomarker measurements are important.