ArticlePloS one2010
DMSO and betaine greatly improve amplification of GC-rich constructs in de novo synthesis.
Article in PloS one, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 59 papers.
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Who cites it
59 citing papers in PubMed.
- Loop-mediated isothermal amplification (LAMP) for quality control of Coscinium fenestratum: a colorimetric platform for detecting Anamirta cocculus adulteration.Protoplasma · 2026Article
- Optimization of PCR amplification and sequencing of GC-rich protein-coding region of CEBPA (CCAAT/enhancer binding protein alpha) in acute myeloid leukemia patients using betaine.Molecular biology reports · 2026Article
- Gungnir codec enabling high error-tolerance and low-redundancy DNA storage through substantial computing power.Nature communications · 2026Article
- Development and Application of a Polymerase Spiral Reaction (PSR)-Based Isothermal Assay for Rapid Detection of Yak (Foods (Basel, Switzerland) · 2025Article
- Antibody-Initiated Loop-Mediated Isothermal Nucleic Acid Amplification (ai-LAMP) as a New Biosensor for Antigen Detection.bioRxiv : the preprint server for biology · 2025Article
- One-Pot Isothermal Nucleic Acid Amplification Assisted CRISPR/Cas Detection Technology: Challenges, Strategies, and Perspectives.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2025Review
- The effects of DMSO on DNA conformations and mechanics.Biophysical journal · 2025Article
- Optimizing PCR amplification of GC-rich nicotinic acetylcholine receptor subunits from invertebrates.Biochemistry and biophysics reports · 2025Article
- Developing, Characterizing, and Modeling CRISPR-Based Point-of-Use Pathogen Diagnostics.ACS synthetic biology · 2025Article
- Innovative nucleic acid detection ofFrontiers in cellular and infection microbiology · 2025Article
- Developing, characterizing and modeling CRISPR-based point-of-use pathogen diagnostics.bioRxiv : the preprint server for biology · 2024Article
- The Rapid Detection ofFoods (Basel, Switzerland) · 2023Article
- Current and Future Advances in the Detection and Surveillance of Biosecurity-Relevant Equine Bacterial Diseases Using Loop-Mediated Isothermal Amplification (LAMP).Animals : an open access journal from MDPI · 2023Review
- Article
- Genetic Authentication of the Medicinal PlantInternational journal of molecular sciences · 2023Article
- Development of subfamily-based consensus PCR assays for the detection of human and animal herpesviruses.European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology · 2023Article
- Engineering highly thermostable Cas12b via de novo structural analyses for one-pot detection of nucleic acids.Cell reports. Medicine · 2023Article
- Maternal plasma and salivary anelloviruses in pregnancy and preterm birth.Frontiers in medicine · 2023Article
- Diagnostic utility and validation of a newly developed real time loop mediated isothermal amplification method for the detection of SARS CoV-2 infection.Journal of clinical virology plus · 2022Article
- Enhancement of a recombinase-aided amplification assay using betaine and pullulan.Infectious medicine · 2022Article
Corrections and comments
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Authors and funding
3 authors.
Funding
Abstract
In Synthetic Biology, de novo synthesis of GC-rich constructs poses a major challenge because of secondary structure formation and mispriming. While there are many web-based tools for codon optimizing difficult regions, no method currently exists that allows for potentially phenotypically important sequence conservation. Therefore, to overcome these limitations in researching GC-rich genes and their non-coding elements, we explored the use of DMSO and betaine in two conventional methods of assembly and amplification. For this study, we compared the polymerase (PCA) and ligase-based (LCR) methods for construction of two GC-rich gene fragments implicated in tumorigenesis, IGF2R and BRAF. Though we found no benefit in employing either DMSO or betaine during the assembly steps, both additives greatly improved target product specificity and yield during PCR amplification. Of the methods tested, LCR assembly proved far superior to PCA, generating a much more stable template to amplify from. We further report that DMSO and betaine are highly compatible with all other reaction components of gene synthesis and do not require any additional protocol modifications. Furthermore, we believe either additive will allow for the production of a wide variety of GC-rich gene constructs without the need for expensive and time-consuming sample extraction and purification prior to downstream application.
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Registered trials
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