Talking Techniques

9/21/2021

The evolution of PCR: From q to dd and beyond

Season 2, Ep. 16
PCR has evolved rapidly over the last few years, an evolution that has been dramatically accelerated by the COVID-19 pandemic. New technologies and approaches have been brought to the fore and the full range of applications for which this technique can be applied have been highlighted, from diagnostics to basic disease research and immunology.This episode explores this evolution, looking at how challenges presented by the pandemic forced the hand of researchers to think outside the box and build on lesser-known PCR technologies. Here, Eddy van Collenburg, Market Development Specialistat Bio-Rad (CA, USA), provides his insight into how ddPCR rose to prevalence during this time and explains how the technology is being used to enable trade, detect new variants and is being used outside of the pandemic.Discover the key role that ddPCR can play in gene therapy research, liquid biopsies, cancer research, single-cell studies and more, in this fascinating overview of the development of PCR and where the technique can go in the future.Contents:Introduction: 00:00-01:30Why was qPCR established as the gold standard for detection?01:30-03:40Limitations of qPCR during the pandemic: 03:40-05:05New varieties of PCR for faster results: 05:05-05:40New varieties for more accurate results: 05:40-06:45ddPCR working principle and advantages: 06:45-09:00Nonclinical ddPCR applications for COVID-19: 09:00-10:45ddPCR in variant detection: 10:45-12:05ddPCR in gene therapy research: 11:05-12:30Liquid biopsy and cancer research: 12:30-15:20Combining ddPCR with NGS in research: 15:20-16:15What are some of the applications of ddPCR that you find most exciting? 16:15-17:10Applications of ddPCR in single-cell studies: 17:10-21:00Improvements in ddPCR: 21:00-21:45Tips for best practice when using ddPCR: 21:45-22:50What would you ask for to improve ddPCR? 21:50-24:20
8/27/2021

Molecular therapeutics: how far have we come and what's on the horizon?

Season 2, Ep. 14
Uncover the history of molecular therapeutics, the staggering current applications and developments of these therapeutics and what their future could hold, in this episode of Talking Techniques.To explore the field of molecular therapeutics and provide an insight into their production, is Aaron Clauson, Product Manager at Zymo research. First, we cover the ins and outs of targeting the therapies and how to select the correct type of molecular therapeutic to begin developing a drug for a specific disease, before looking at some of the setbacks and safety errors that have occurred during the development of molecular therapeutics and how researchers have learned from these tragic events.Aaron also highlights the vital role of plasmids in almost all molecular therapeutic development, detailing the importance of keeping plasmid solutions and transfections free of endotoxins, revealing key solutions for avoiding these contaminants.Listen today to get all you need to know about molecular therapeutics, from their inception to their most exciting examples and how we can better communicate their benefits to the public in order to avoid mistrust and confusion - as has occurred during the COVID-19 pandemic.Contents:What counts as a molecular therapeutic? 01:45-02:40Well known molecular therapeutics: 02:40-05:00What can molecular therapeutics achieve that synthetic chemicals can’t? 05:00-05:55Targeting molecular therapeutics and picking the right type of therapeutic: 05:55-08:25Selecting the appropriate molecular therapeutic: 08:35-11:00Setbacks and safety concerns in gene therapies: 11:00-15:45Preventing off-target effects in gene therapies: 15:45-17:35Challenges in the development of molecular therapeutics: 17:35-19:05Plasmids in molecular therapeutic development: 19:05-23:55Avoiding endotoxins: 23:55-27:45What fields do you find most exciting? 27:45-28:50Better communication of gene therapies to the public: 28:50-32:00 Improving molecular therapeutic development: 32:00-37:10
8/17/2021

Adeno-associated viral therapies: harnessing the power of HPLC

Season 2, Ep. 13
This episode centers on the critical role that high-performance liquid chromatography (HPLC) plays in the production of adeno-associated viral (AAV) therapies, the latest advances in AAV design and the next steps in their development.Filling me in on all things HPLC and AAVs is Bella Neufeld, Director of R&D at Teknova. Bella reveals the essential validation and quality control steps required for the production of successful and safe AAV therapies and the role that both HPLC and liquid chromatography–mass spectrometry (LCMS) play in these steps. We also explore the latest development targets for these therapies, looking at how AAVs can be targeted to specific sites and tissues before descending into the treacherous topic of reproducibility in the field.Listen today to find out the latest tips for best practice when working with HPLC, LCMS the advantages of LCMS over other analytical techniques to assess empty and full capsids and the key principles of AAV production!ContentsIntro: 00:00-01:25Explaining HPLC and its benefits: 01:25-03:30Developing adeno-associated viruses: 03:30-05:25Chromatography in validation vs production of AAVs: 05:25-06:15Quality control vs hands on time: 06:15-07:35Challenges in AAV production: 07:35-08:55Examining the level of empty and full capsids: 08:55-10:00The role of LCMS in determining and validating AAV contents: 10:00-11:30Tips for best practice when using HPLC: 12:30-13:10The future of HPLC: 13:10-14:20Latest advances in AAV development: 14:20-15:50Targeting AAVs: 15:50-17:20Reproducibility issues in AAV development: 17:20-19:35Outro: 19:35-21:10
8/16/2021

Organoids: advancing drug discovery and cancer research

Season 2, Ep. 12
As the transition from 2D to 3D cell cultures, or organoids, as the gold standard for modeling basic biology and disease continues, these models are being utilized in ever more intricate and impactful ways.This episode covers the rise of organoids and their advantages over 2D cell cultures, before exploring just how representative these models are becoming and their new applications in drug discovery. To find out all this and more, I speak to Oksana Sirenko, Senior Scientist at Molecular Devices, who works extensively with the company's organoid model development programs.Oksana reveals the potential of these models in cancer therapeutics, driven forward by new patient-derived organoids, and notes the challenges presented by imaging these thicker, 3D models. We also explore the potential discoveries that can be unlocked by combining organoids with organ-on-a-chip models.Contents:What are organoids? 02:10-03:20Functions replicated by organoids? 3:20-04:00What are the advantages of 3D over 2D cell cultures? 04:00-05:40Just how representative are these models? 04:40-07:10Intestinal organoid formation 07:10-08:50The most exciting application of organoids 08:50-10:10Organoids in drug development 10:10-11:20Methodology of organoids in drug screening studies 11:20-13:30Examples of organoids in cancer drug discovery: 13:30-16:00Limitations of patient-derived and regular organoid models: 16:00-17:50Body-on-a-chip concepts and examples: 17:50-22:00Challenges posed by organoids for drug discovery: 22:00-24:20Techniques improving organoid models and analysis: 24:20-25:50Imaging organoid models: 25:50-26:55Tips for best practice when working with organoids: 26:55-28:20Organoid method development: 28:20-31:10One thing to improve organoid models or analysis for drug screening: 31:10-33:07To find out more about the research and solutions discussed in this episode visit: www.moleculardevices.com/TechTalk8.
6/25/2021

The ethics of whole-genome sequencing: from race and religion to science fiction

Season 2, Ep. 11
Welcome to part two of this two-part episode on whole-genome sequencing (WGS) in rare disease research and treatment. In part one we discussed the impacts the technology has had on both improving the clinical diagnosis and management of rare diseases and on drug development and basic research studies.This episode goes beyond the technology’s capabilities and into the debates surrounding how, when and why they should be used. What needs to change for the technique to be applied equitably? And how does reality stack up against science fiction in terms of people’s perceptions of ubiquitous WGS?To do this I speak with David Dimmock, Senior Medical Director of the Rady Children’s Institute for Genomic Medicine (CA, USA); Christine Stanley, Chief Director of Clinical Genomics, Variantyx (MA, USA); and Take Ogawa, Vice President of Sales & Marketing at Psomagen (MD, USA). Each of these speakers provides their opinions on the impacts of race, religion and way of life on the application of WGS and how much is too much data.Contents:Introductions: 00:00-02:20Gauging public perceptions of routine WGS 02:20-05:40The role of industry in improving WGS accessibility and capabilities: 05:40-07:35The appropriate use of pre-symptomatic and predictive risk findings: 08:00-11:15Debating the discovery of untreatable diseases: 11:15-12:45The impact of data biases in gene sequencing and the lack of gene references for different populations: 12:45-14:45The impact of religion and lifestyle on ethical considerations in the use of WGS data: 14:45-16:00Dealing with unintended findings or results that impact relatives: 16:00-16:45The value of WGS in pharmacogenomic variant findings and challenging the ACMG59 list: 16:45-19:20Striking the balance between fully utilizing WGS for all potential findings and ensuring maximum participation for all groups: 19:20-22:45Filtering data to avoid exclusion: 22:45-24:10Who owns WGS data? 24:10-25:15Consent conversations: 25:15-26:30Final comments: 26:30-27:15
6/18/2021

From space to the streets: infectious disease sampling in extreme conditions and extraordinary circumstances

Season 2, Ep. 10
In this episode, we take a look at one of the key steps of infectious disease research, sample collection; exploring the challenges associated with the step and the solutions available to improve infectious disease research around the globe.Exploring the trials and tribulations of infectious disease sample collection with me is Stanislav Forman, Product Developer at Zymo Research (CA, USA). Stan explains the extreme environments in which sample collection often occurs; revealing the difficult transit and storage requirements –often involving cold chain logistics –that can derail infectious disease studies early on.Find out how key solutions to these problems have been successfully workshopped with the US military and have even been used in work with NASA to conduct safe sample collection in space. And discover just how successful these solutions can be when faced with the blazing wreckage of a transit vehicle…ContentsIntroductions: 00:00-01:20Key challenges of sample collection for infectious disease research: 01:20-02:30Additional challenges presented by COVID-19: 02:30-05:00The emergence of at-home testing: 05:00-07:00Established practices for sample collection and introducing DNA/RNA shield: 07:00-09:00Protecting DNA/RNA samples: 09:00-11:00Working with NASA on sample collection and protection in space: 11:00-14:20Protecting samples in a blazing transit van: 14:20-15:25The impact of inactivating and preserving sample collection products on infectious disease research: 15:30-16:30Areas for improvement in sample collection: 16:30-17:30Closing statements: 17:30-19:20
6/11/2021

Revealing the regulome: using multiomic approaches to explore epigenetics and DNA expression

Season 2, Ep. 9
Explore the realm of epigenetics and gene expression, discovering the different omic ‘lenses’ used to examine these processes and the power of single-cell studies to reveal a comprehensive multiomic view of cells and their biology.This episode features the ever-buoyant Will Greenleaf, Associate Professor of Genetics at Stanford University School of Medicine (CA, USA) and head of the Greenleaf Lab. Will takes us through his lab’s work, detailing his attempts to understand the compacting of DNA into cell nuclei and how this impacts gene expression and epigenetic regulation.Discover the emerging new research focus that Will has coined the “Regulome” – the collection of physical molecular components that control gene expression through factors such as chromatin accessibility. To explore these new frontiers Will often encounters questions with no technologies available to ask them. So what does he do? Invent the technologies he needs! Find out how his lab came to develop the, now widely used, ATAC-Seq and learn of the new tools that he is looking to develop to solve even more perceptive questions.This episode has it all, and also reveals the potential of Will’s single-cell multiomic approaches in cancer research, virology and HIV. Contents:·Introductions: 00:00-02:00·The exploring the physical genome: 02:00-03:15·Chromatin accessibility: 03:15-05:15·Introducing the ‘Regulome’: 05:15-07:15·The importance of multiomics for the study of gene expression: 07:15-07:55·DNA methylation and histone organization: 07:55-10:30·Real-world impact of this research: 10:30-12:30·Will’s varied career path: 12:30-16:15·Combining multiomic datasets with computational biology and physics experience: 16:15-18:40·Single-cell techniques: 18:40-21:50·Developing ATAC-Seq: 21:50-23:50·New tools in development: 23:50-27:30·Insights into the epigenetics of cancer: 27:30-29:35·Insights into virology and HIV latency: 29:35-32:10·One wish to help explore the regulome. 32:10-33:00
5/28/2021

The oncolytic Trojan horse: Immune cells, the tumor microenvironment and the invasion of neurons

Season 2, Ep. 8
This episode, supported by Bethyl, delves into the realm of the tumor microenvironment (TME), exploring the cells that reside there and how they interact to promote tumor growth and metastasis. Discover how immune cells are attracted and manipulated by tumor cells enabling the cancer to invade neurons where they can then travel throughout the body.Providing an expert's insight into this topic is Moran Amit, Assistant Professor at the MD Anderson Cancer Center (TX, USA). Moran exposes the nebulous interactions in the TME and describes the techniques that he uses to interrogate it, in the hope that by further understanding these interactions we will be able to design more successful, targeted therapies for neurotropic cancers.Contents:·Introductions: 00:00-01:20·The key cells of the TME: 01:20-02:50·How cells of the TME support the tumor: 02:50-05:45·How cancer cells influence and impact immune cells: 05:45-07:25·The invasion of cancers into the neurons: 07:27-9:45·Recruitment of neurons to the TME: 09:45-10:55·The evolution of neuron recruitment to the TME: 10:55-15:00·Techniques involved in the study of the TME: 15:00-17:00·Tips for best practice: 17:00-18:00·Tumors with neural networks: 18:00-20:05·Clinical impact of research: 20:05-21:50·The cancers impacted the most by this research: 21:50-23:25·The impact of neural invasion on patients and neurons: 23:25-25:30·One thing to improve our understanding of the TME: 25:30-28:00