ArticleBioTechniques2018
Limitations of methods for measuring the concentration of human genomic DNA and oligonucleotide samples.
Article in BioTechniques, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed, 29 citations in OpenAlex.
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- Fluorescent Techniques for RNA Detection in Nanoparticles.Methods in molecular biology (Clifton, N.J.) · 2024Article
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- Assessment and validation of enrichment and target capture approaches to improveJournal of clinical microbiology · 2023Article
- Calibration of cell-free DNA measurements by next-generation sequencing.American journal of clinical pathology · 2023Article
- Quantitation of Residual Host Cell DNA in Recombinant Adeno-Associated Virus Using Droplet Digital Polymerase Chain Reaction.Human gene therapy · 2023Article
- Production and Characterization of a SARS-CoV-2 Nucleocapsid Protein Reference Material.ACS measurement science au · 2022Article
- Characterization of a SARS-CoV-2 spike protein reference material.Analytical and bioanalytical chemistry · 2022Article
- Quantification of DNA through the NanoDrop Spectrophotometer: Methodological Validation Using Standard Reference Material and Sprague Dawley Rat and Human DNA.International journal of analytical chemistry · 2020Article
- Absolute Quantification of RNA or DNA Using Acid Hydrolysis and Mass Spectrometry.Analytical chemistry · 2019Article
- Development and interlaboratory evaluation of a NIST Reference Material RM 8366 for EGFR and MET gene copy number measurements.Clinical chemistry and laboratory medicine · 2019Article
- Multilaboratory Assessment of a New Reference Material for Quality Assurance of Cell-Free Tumor DNA Measurements.The Journal of molecular diagnostics : JMD · 2019Article
Corrections and comments
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
Abstract
We compared different methods (absorbance, fluorescent dye-binding, and digital PCR) for measuring the concentrations of human genomic DNA from cultured cells and absorbance measurements of a synthetic DNA oligonucleotide. NIST Standard Reference Material (SRM) 2082, a pathlength absorbance standard, was used to benchmark the absorbance measurements done with microvolume spectrophotometers and a microvolume plate reader. Control absorbance values were measured on a high accuracy spectrophotometer and a NIST calibrated pathlength cuvette. Measurements of the human genomic DNA sample were done with several types of fluorescent dye binding assays using different DNA calibrators. The fluorescent dye binding methods gave different results for genomic DNA depending upon the type of DNA calibrator and the fluorescent dye that was used. The human genomic DNA sample was also characterized by using six different droplet digital PCR assays (amplicons located on different chromosomes) to measure the average copy number. Conversion of the digital PCR data to copy numbers was sensitive to the droplet size used for calculations and conversion to mass concentration was dependent upon the molecular weight of the human genome used for the calculations. The results from the different methods were compared and the caveats for each measurement method were discussed.
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Registered trials
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