ArticleBMC biotechnology2003
Performance of high-throughput DNA quantification methods.
Article in BMC biotechnology, 2003. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 25 papers.
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Who cites it
25 citing papers in PubMed, 114 citations in OpenAlex.
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- Comparison of Automated and Manual DNA Isolation Methods of Liquid-Based Cytology Samples.Biopreservation and biobanking · 2019Article
- Surface Modification of Silicon Pillar Arrays To Enhance Fluorescence Detection of Uranium and DNA.ACS omega · 2017Article
- Comparison of DNA quantification methodology used in the DNA extraction protocol for the UK Biobank cohort.BMC genomics · 2017Article
- A Biospecimen Proficiency Testing Program for Biobank Accreditation: Four Years of Experience.Biopreservation and biobanking · 2016Article
- A comparison between quantitative PCR and droplet digital PCR technologies for circulating microRNA quantification in human lung cancer.BMC biotechnology · 2016Article
- Variation in pre-PCR processing of FFPE samples leads to discrepancies in BRAF and EGFR mutation detection: a diagnostic RING trial.Journal of clinical pathology · 2015Article
- Considerations for accurate gene expression measurement by reverse transcription quantitative PCR when analysing clinical samples.Analytical and bioanalytical chemistry · 2014Review
- In vitro mesenchymal trilineage differentiation and extracellular matrix production by adipose and bone marrow derived adult equine multipotent stromal cells on a collagen scaffold.Stem cell reviews and reports · 2013Article
- Evaluation of digital PCR for absolute RNA quantification.PloS one · 2013Article
- DNA qualification workflow for next generation sequencing of histopathological samples.PloS one · 2013Article
- Collection of human genomic DNA from neonates: a comparison between umbilical cord blood and buccal swabs.American journal of obstetrics and gynecology · 2011Article
- Detection of somatic mutations by high-resolution DNA melting (HRM) analysis in multiple cancers.PloS one · 2011Article
Corrections and comments
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Authors and funding
6 authors at 2 institutions in 1 country.
Funding
Abstract
backgroundThe accuracy and precision of estimates of DNA concentration are critical factors for efficient use of DNA samples in high-throughput genotype and sequence analyses. We evaluated the performance of spectrophotometric (OD) DNA quantification, and compared it to two fluorometric quantification methods, the PicoGreen assay (PG), and a novel real-time quantitative genomic PCR assay (QG) specific to a region at the human BRCA1 locus. Twenty-Two lymphoblastoid cell line DNA samples with an initial concentration of approximately 350 ng/uL were diluted to 20 ng/uL. DNA concentration was estimated by OD and further diluted to 5 ng/uL. The concentrations of multiple aliquots of the final dilution were measured by the OD, QG and PG methods. The effects of manual and robotic laboratory sample handling procedures on the estimates of DNA concentration were assessed using variance components analyses.
resultsThe OD method was the DNA quantification method most concordant with the reference sample among the three methods evaluated. A large fraction of the total variance for all three methods (36.0-95.7%) was explained by sample-to-sample variation, whereas the amount of variance attributable to sample handling was small (0.8-17.5%). Residual error (3.2-59.4%), corresponding to un-modelled factors, contributed a greater extent to the total variation than the sample handling procedures.
conclusionThe application of a specific DNA quantification method to a particular molecular genetic laboratory protocol must take into account the accuracy and precision of the specific method, as well as the requirements of the experimental workflow with respect to sample volumes and throughput. While OD was the most concordant and precise DNA quantification method in this study, the information provided by the quantitative PCR assay regarding the suitability of DNA samples for PCR may be an essential factor for some protocols, despite the decreased concordance and precision of this method.
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