ArticleScientific reports2020
Two-phase wash to solve the ubiquitous contaminant-carryover problem in commercial nucleic-acid extraction kits.
Article in Scientific reports, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
What it found
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
17 citing papers in PubMed, 22 citations in OpenAlex.
- Development of a Direct Cell-to-PCR Lysis Buffer Using Optimized Non-Ionic Detergents for RNA-Extraction-Free RT-qPCR.Methods and protocols · 2026Article
- Revolutionizing Healthcare With Paper-Based Nucleic Acid Testing.Exploration (Beijing, China) · 2026Review
- DNA sequence contamination analyzer (DNASCAN): a supervised analysis toolkit for detecting and removing DNA contaminants.BMC bioinformatics · 2026Article
- Optimizing Lysis and Extraction Workflows for Enrichment-Free qPCR Detection ofPathogens (Basel, Switzerland) · 2026Article
- Comparative Evaluation of Automated Nucleic Acid Extraction Systems for DNA and RNA Viral Target.Pathogens (Basel, Switzerland) · 2026Article
- The impact of DNA extraction methods on species quantification and apparent community composition of in vitro oral biofilms.FEMS microbiology letters · 2026Article
- Optimization and validation of the real-time polymerase chain reaction protocol for porcine gelatin detection in processed food and cosmetic products.Open veterinary journal · 2026Article
- Single-Tube Capture, Concentration, and Genomic Extraction of a Human Norovirus Surrogate Using Magnetic Ionic Liquids.Analytical chemistry · 2025Article
- Skin biopsy processing for rapid molecular diagnosis and histopathologic interpretation: application to Kaposi sarcoma in East Africa.Infectious agents and cancer · 2025Article
- An inhibitor-free, versatile, fast, and cheap precipitation-based DNA purification method.PloS one · 2025Article
- Method for lysis and paper-based elution-free DNA extraction with colourimetric isothermal amplification.Scientific reports · 2024Article
- Impact of relative humidity on SARS-CoV-2 RNA extraction using Nextractor automated extraction system.Journal of biological methods · 2024Article
- Chemical Trends in Sample Preparation for Nucleic Acid Amplification Testing (NAAT): A Review.Biosensors · 2023Review
- An evaluation of DNA extraction methods on historical and roadkill mammalian specimen.Scientific reports · 2023Article
- Filtration-assisted magnetofluidic cartridge platform for HIV RNA detection from blood.Lab on a chip · 2022Article
- Improved DNA extraction on bamboo paper and cotton is tightly correlated with their crystallinity and hygroscopicity.PloS one · 2022Article
- Two-phase wash to solve the ubiquitous contaminant-carryover problem in commercial nucleic-acid extraction kits.Scientific reports · 2020Article
Corrections and comments
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
Abstract
The success of fundamental and applied nucleic acid (NA) research depends on NA purity, but obtaining pure NAs from raw, unprocessed samples is challenging. Purification using solid-phase NA extractions utilizes sequential additions of lysis and wash buffers followed by elution. The resulting eluent contains NAs and carryover of extraction buffers. Typically, these inhibitory buffers are heavily diluted by the reaction mix (e.g., 10x dilution is 1 µL eluent in 9 µL reaction mix), but in applications requiring high sensitivity (e.g., single-cell sequencing, pathogen diagnostics) it is desirable to use low dilutions (e.g., 2x) to maximize NA concentration. Here, we demonstrate pervasive carryover of inhibitory buffers into eluent when several commercial sample-preparation kits are used following manufacturer protocols. At low eluent dilution (2-2.5x) we observed significant reaction inhibition of polymerase chain reaction (PCR), loop-mediated isothermal amplification (LAMP), and reverse transcription (RT). We developed a two-phase wash (TPW) method by adding a wash buffer with low water solubility prior to the elution step. The TPW reduces carryover of extraction buffers, phase-separates from the eluent, and does not reduce NA yield (measured by digital PCR). We validated the TPW for silica columns and magnetic beads by demonstrating significant improvements in performance and reproducibility of qPCR, LAMP, and RT reactions.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.