Boosting PCR accuracy – tips for maximizing amplification fidelity

In this Mol Bio Minutes episode, Laurynas Alijošius shares a personal story that every molecular biologist can relate to—running PCR, cloning, and sequencing, only to discover frustrating errors in the DNA. This episode dives deep into PCR accuracy and why it matters for everything from sequencing to cloning and long-read library prep. Laurynas breaks down the major contributors to PCR error, including the fidelity of DNA polymerase, primer design flaws, template impurities, and suboptimal cycling conditions. He then offers a range of solutions—like switching to high-fidelity enzymes, using ready-to-go master mixes, and optimizing magnesium ion concentrations. He also explains how reducing cycle numbers and fine-tuning annealing temperatures can minimize unwanted amplification and ensure more reliable data.Whether you're troubleshooting or proactively optimizing your workflow, this episode is packed with tips and tools to help you increase PCR accuracy, reduce costs, and save time. Episode notes contain links to enzyme comparisons, primer design tools, and cycling guides to help you PCR with precision.Helpful resource links mentioned in this episode:Thermo Scientific web tools for primer design and analysis, and moreThe PCR Learning Center with lots of helpful tips and informationLearn more about PCR reagents and enzymesBrowse and purchase PCR enzymes and master mixesAccess the PCR troubleshooting guideDownload the molecular biology handbook  Subscribe to get future episodes as they drop and if you like what you’re hearing we hope you’ll share a review or recommend the series to a colleague.  Visit the Invitrogen School of Molecular Biology to access helpful molecular biology resources and educational content, and please share this resource with anyone you know working in molecular biology.  For Research Use Only. Not for use in diagnostic procedures.

In this episode of Speaking of Mol Bio, Dr. Cath Moore of the Australian Genome Research Facility (AGRF) discusses how molecular biology technologies are helping to shape Australia’s scientific landscape—from clinical genomics and conservation to bioremediation and agriculture. With over 20 years of experience in both academia and industry, Dr. Moore reflects on the remarkable evolution of genomic tools, from Sanger sequencing to high-resolution spatial multiomics.She unpacks AGRF’s mission to democratize access to emerging technologies and highlights its role as an early adopter of platforms that help scientists translate academic research into real-world impact. Topics include non-mass spec proteomics, mine site rehabilitation through soil microbiome analysis, and the role of systems biology in modern science.Dr. Moore also discusses the importance of community education and literacy around genomics, emphasizing how public understanding is key to the safe adoption of emerging technologies like synthetic biology. Finally, she shares career insights and advice for aspiring scientists: stay curious, stay broad, and don’t be afraid to pivot when your work no longer brings joy. Subscribe to get future episodes as they drop and if you like what you’re hearing we hope you’ll share a review or recommend the series to a colleague.  Visit the Invitrogen School of Molecular Biology to access helpful molecular biology resources and educational content, and please share this resource with anyone you know working in molecular biology.  For Research Use Only. Not for use in diagnostic procedures.

In this Mol Bio Minutes episode, Laurynas Alijošius breaks down how to run fast PCR to save time and increase lab efficiency. He explains how to choose the right thermal cycler with fast ramp speeds, select low-volume and thin-walled PCR plastics, and use engineered DNA polymerases that offer rapid elongation and hot-start capability. Laurynas also covers practical tips for optimizing reaction components, shortening cycling protocols, and reducing waste. Whether you’re aiming to finish your experiment before dinner or streamline your workflow long-term, this episode delivers everything you need to master the art of fast PCR.Helpful resource links mentioned in this episode:Molecular biology handbook – An extensive resource for all things molecular biologyApplied Biosystems thermal cyclers – Including those that support fast PCRPCR consumables – Four key attributes to considerPCR plastics selection tool – Find the right plastics for your instrument and fast PCRDNA polymerases – Four key characteristics to know and considerPCR setup optimization – Six critical things to optimize for optimal PCR resultsEnzymes and master mixes – Get the right reagents to drive your PCR reactionCycling optimization – Instrument considerations for PCR success Subscribe to get future episodes as they drop and if you like what you’re hearing we hope you’ll share a review or recommend the series to a colleague.  Visit the Invitrogen School of Molecular Biology to access helpful molecular biology resources and educational content, and please share this resource with anyone you know working in molecular biology.  For Research Use Only. Not for use in diagnostic procedures.

This episode teaches that innovation is born at the intersection of curiosity and persistence. Dr. Gudrun Stengel, co-founder and CEO of Alida Biosciences, reveals how her startup is transforming the landscape of RNA research through a novel technology called proximity barcoding. Dr. Stengel’s story exemplifies the power of entrepreneurial spirit in driving scientific discovery, offering a glimpse into how one idea can reshape an entire field.At Alida Biosciences, Dr. Stengel and her team are pioneering new tools for detecting RNA modifications, a largely unexplored realm of epigenetics. Using their proximity barcoding platform, researchers can read multiple RNA modifications simultaneously, uncovering potential biomarkers and therapeutic targets for diseases like cancer, Alzheimer’s, and diabetes. This technology bridges a critical gap in multiomics, allowing scientists to dive deeper into how epigenetic changes influence gene expression and cellular behavior.Beyond the lab, Dr. Stengel shares her experience as a first-time founder, balancing scientific rigor with startup life. From managing a team to fundraising, her journey underscores the importance of resilience, optimism, and maintaining a long-term vision. She also offers valuable advice for budding scientists, encouraging them to embrace challenges and remain persistent in the face of setbacks. Subscribe to get future episodes as they drop and if you like what you’re hearing we hope you’ll share a review or recommend the series to a colleague.  Visit the Invitrogen School of Molecular Biology to access helpful molecular biology resources and educational content, and please share this resource with anyone you know working in molecular biology.  For Research Use Only. Not for use in diagnostic procedures.

Sustainability in the lab isn't just a trend—it's a responsibility. In this quick but powerful Mol Bio Minutes episode, sustainability expert Sune Lund Sporring shares actionable tips to reduce energy use, manage lab waste, and transition to greener materials like Thermo Fisher’s Sustain Series bio-based plastics.Discover the difference between bio-based and biodegradable, why second-generation feedstocks are a game-changer, and how to make low-impact changes without compromising performance. Learn how small shifts—like using aluminum beads instead of water in a bath or switching to carbon-reduced plastics—can significantly cut your lab’s carbon footprint. And remember: if greener options aren’t available, your demand can help shape the market.Helpful resource links mentioned in this episode:My Green Lab – Facts and resources about lab sustainabilityEnergy use of fume hoods – Energy use and savings ideas for fume hoodsGreener by Design – Thermo Fisher’s approach to green solutionsLab Armor™ Beads – A sustainable option to replace water in water bathsInstrument trade up program – trade in your equipment to be green and give it a second lifeLab plastic waste – Stats about plastic waste in labsDoing something about it – Thermo Fisher’s lower carbon plastics solutionSustain Series PCR plastics – Lower carbon footprint, without workflow interruptionReasons to believe – Fact sheet about Sustain Series PCR plastics Subscribe to get future episodes as they drop and if you like what you’re hearing we hope you’ll share a review or recommend the series to a colleague.  Visit the Invitrogen School of Molecular Biology to access helpful molecular biology resources and educational content, and please share this resource with anyone you know working in molecular biology.  For Research Use Only. Not for use in diagnostic procedures.

Join us for an insightful conversation with Dr. Nadav Ahituv, a leader in human genetics and gene regulation at UCSF. He shares his personal journey from scoliosis patient to genetic researcher, exploring the mysteries of non-coding DNA, massively parallel reporter assays, and CRISPR-based therapeutic innovations.We dive into his lab’s diverse projects, from decoding bat wing development and diet adaptation to tackling complex diseases like scoliosis and cancer through gene modulation. Learn how cloning is used alongside technologies like AI, genome-wide sequencing, and CRISPR to revolutionize our understanding of regulatory elements and shape the future of precision medicine.Plus, discover how a surprising approach—using modified fat cells—could be a game-changer in cancer therapy. This episode is a must-listen for anyone fascinated by the intersection of genetics, technology, and evolution. Subscribe to get future episodes as they drop and if you like what you’re hearing we hope you’ll share a review or recommend the series to a colleague.  Visit the Invitrogen School of Molecular Biology to access helpful molecular biology resources and educational content, and please share this resource with anyone you know working in molecular biology.  For Research Use Only. Not for use in diagnostic procedures.

With prior Mol Bio Minutes episodes covering DNA form migration and staining considerations for nucleic acid gel electrophoresis, we tie it all together with this great set of overall tips, tricks and resources for the topic. Anyone that’s ever run a gel has undoubtedly produced gels with smeared, faint or poorly separated bands. What causes these and how can you avoid them?  Well, Aistė Polikaitytė, Scientist at Thermo Fisher Scientific is here to cover the likely causes and troubleshooting tips to help avoid the most common gel issues. She touches on how much sample to load, the importance of reagent selection, gel preparation, separation conditions, staining, as well as purification and contamination considerations. Helpful resource links mentioned in this episode:Selection guide for electrophoresis dyes and buffersLearn how using precast E-Gel agarose gel can help avoid common issuesA helpful troubleshooting guide for nucleic acid gel electrophoresisView an on-demand webinar covering these topics in more detail Subscribe to get future episodes as they drop and if you like what you’re hearing we hope you’ll share a review or recommend the series to a colleague.  Visit the Invitrogen School of Molecular Biology to access helpful molecular biology resources and educational content, and please share this resource with anyone you know working in molecular biology.  For Research Use Only. Not for use in diagnostic procedures.

It could be argued that biology has always boiled down to chemistry, and that chemistry has always boiled down to physics. However, not many would deny that the fields of biology and chemistry are overlapping more than ever, with both leveraging computing methods, also more than ever. This conversation with Dr. Ramesh Jha, Technical Staff Member at Los Alamos National Laboratory (LANL), crosses biology, chemistry, and computing methods. The work of his biome team at LANL uses computational tools to inform the design of enzymes that are produced via PCR-based cloning and then expressed in microbes. They use fluorescent gene circuits in these microbes, along with flow cytometry, to screen these large libraries for advantageous gain-of-function variants. When they find an interesting mutation, they isolate it, sequence it, and produce and evaluate those biocatalytic enzymes for bioremediation, biomanufacturing, and other important applications. Ramesh makes this complex and interdisciplinary science approachable and gives hope to how it could help address problems of “forever chemicals” and other environmental and manufacturing challenges. Join us for this interesting and inspiring conversation.  Subscribe to get future episodes as they drop and if you like what you’re hearing we hope you’ll share a review or recommend the series to a colleague.  Visit the Invitrogen School of Molecular Biology to access helpful molecular biology resources and educational content, and please share this resource with anyone you know working in molecular biology.  For Research Use Only. Not for use in diagnostic procedures.

You can run the perfect agarose gel to separate your nucleic acid fragments but if you don’t stain and image the gel properly, it’s all for not. In this second installment of Mol Bio Minutes we take a look at the staining considerations for nucleic acid gel electrophoresis with Paulius Palaima, Product Manager at Thermo Fisher Scientific. He covers the range of stains and staining approaches available while calling out pros, cons and considerations for each. How these recommendations change, depending on your sample, is also covered in this approachable but informative episode. Helpful resource links mentioned in this episode:A helpful DNA stain selection guideRNA stain options and detailsEffects of dyes on gel electrophoresis properties Subscribe to get future episodes as they drop and if you like what you’re hearing we hope you’ll share a review or recommend the series to a colleague.  Visit the Invitrogen School of Molecular Biology to access helpful molecular biology resources and educational content, and please share this resource with anyone you know working in molecular biology.  For Research Use Only. Not for use in diagnostic procedures.

Topics and terms such as biosafety, biosecurity, containment, and contamination are things most of us have heard of and think about at some level, but with the pace of molecular biology moving faster than ever, these are topics with implications that are reaching farther than ever. We’re joined by Dr. Ryan Burnette and Dr. Lauren Richardson from Merrick and company for this episode, and they’re ace communicators that help walk us through the expanding horizon and implications of these topics.This conversation starts on the basic topics, like what biocontainment is and what’s needed for each of the four levels of biosafety labs, but it quickly moves beyond, shining a light on the security and containment needs for more than just the organisms. We hear about how the data and methods used to do modern molecular biology, as well as the data generated in the experiments, are equally precious and in need of protection and containment. With public health and safety on the line, and an acknowledgement that the pace of science moves faster than that of policy, we get into the idea of who really owns responsibility for protecting data. Your role might be more important than you know, so don’t miss this conversation that will make you pause and think! Subscribe to get future episodes as they drop and if you like what you’re hearing we hope you’ll share a review or recommend the series to a colleague.  Visit the Invitrogen School of Molecular Biology to access helpful molecular biology resources and educational content, and please share this resource with anyone you know working in molecular biology.  For Research Use Only. Not for use in diagnostic procedures.