Cell-free Dna Quantification
Cell-free Dna Quantification - Related Videos
07:53
Quantification of Colonic Stem Cell Mutations
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Cited by 1 •
2015
We report significant improvements for the reproducible measurement of somatic colonic stem cell mutations after exposure of mice to potential DNA damaging agents.
10:47
High-Throughput Image-Based Quantification of Mitochondrial DNA Synthesis and Distribution
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2023
A procedure for studying the dynamics of mitochondrial DNA (mtDNA) metabolism in cells using a multi-well plate format and automated immunofluorescence imaging to detect and quantify mtDNA synthesis and distribution is described. This can be further used to investigate the effects of various inhibitors, cellular stresses, and gene silencing on mtDNA metabolism.
08:03
Simultaneous Assessment of Cardiomyocyte DNA Synthesis and Ploidy: A Method to Assist Quantification of Cardiomyocyte Regeneration and Turnover
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Cited by 25 •
2016
Quantification of cardiomyocyte turnover is challenging. The protocol described here makes an important contribution to this challenge by enabling accurate and sensitive quantification of neo-cardiomyocyte nuclei generation and nuclei ploidy.
11:37
Simultaneous DNA-RNA Extraction from Coastal Sediments and Quantification of 16S rRNA Genes and Transcripts by Real-time PCR
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Cited by 24 •
2016
A protocol to quantify bacterial 16S rRNA genes and transcripts from coastal sediments via real-time PCR is provided. The methodology includes the co-extraction of DNA and RNA; preparation of DNA-free RNA; and 16S rRNA gene and transcript quantification via RT-q-PCR, including standard curve construction.
07:37
Circulating MicroRNA Quantification Using DNA-binding Dye Chemistry and Droplet Digital PCR
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Cited by 10 •
2016
A sensitive and accurate method for cell-free microRNAs quantification using a dye-based chemistry and droplet digital PCR technology is described.
FAQs
Interpretation depends on what the measured DNA represents. Total cell-free DNA can reflect tissue injury or immune activity, whereas a sequence-specific signal can indicate DNA associated with a microbial target. Because these measurements address different biological sources, researchers should avoid treating total abundance as a direct substitute for evidence of infection. This distinction guides biomarker selection.
Fluorescence-based assays and amplification-based assays answer related but different questions. Fluorescence provides a measurement of DNA in the extracted sample, while quantitative PCR or digital PCR measures a selected sequence and can therefore focus on pathogen-specific or other defined targets. The choice determines whether the result reflects broad DNA abundance or a targeted molecular signal.
Targeted measurement links the assay to a defined microbial sequence rather than to extracellular DNA as a whole. That specificity can help investigators assess microbial presence, while total cell-free DNA provides complementary information about host damage or immune activity. Using both readouts may help separate infection-related signals from broader changes in the host response.
Repeated measurements can show how extracellular DNA changes over time during an infection or immune response. A total-DNA trend may track changing host injury or immune activity, while a pathogen-specific trend may provide information about microbial presence. In treatment-monitoring studies, comparing measurements at multiple time points can connect molecular changes with the course of disease or response.
Cell-free DNA quantification generally begins with collection of plasma, serum, or another biological fluid, followed by DNA isolation. The recovered material is then measured either with a fluorescence-based assay or with sequence-specific amplification using quantitative PCR or digital PCR. This workflow separates sample preparation from measurement and allows investigators to select total or targeted DNA readouts.
It provides a molecular readout for examining how immune activity and tissue injury alter extracellular nucleic acids. Investigators can measure total cell-free DNA to study broad host-associated changes, then use sequence-specific assays when infection-related targets are important. These data support biomarker development and help relate immune responses to measurable changes in biological fluids.