Talking Techniques


How PCR has prevailed during the COVID-19 pandemic

Season 2, Ep. 5
In Part 3 of our COVID-19 diagnostics and detection miniseries, supported byRoche, we explore the contributionPCRhas made towards diagnosis during the pandemic. Providing me with an insight into the world of PCR diagnostics is Tyler Miller, Clinical Pathology Resident and Research Fellow atMassachusetts General Hospital(MA, USA), where he was instrumental in setting up the testing regimen for the Hospital.Ty details the attributes of PCR that lead to it becoming the gold standard for diagnostic tests, before explaining the variance in clinical detection rate during a patient’s disease course and how this variation correlates with the infectivity of the patient. Ty also delves into the variety of sample collection methods available, how each of these methods compares in terms of sensitivity and their ability to be integrated into exciting novel PCR techniques.We also look at the work of theBroad Institutein establishing a mass testing effort that involved automation, workflow optimization and hundreds of new staff, ultimately leading to the delivery of 100,000 tests a day, almost 5% of the USA’s total COVID-19 testing at the time. All of this rapid work has led to dramatic changes in the PCR technique and process. These changes are perhaps exemplified by PCR testing without RNA extraction and purification, which Ty explains was partly developed due to the limited supplies available to researchers at the beginning of the pandemic.ContentsIntroduction: 00:00-02:00Why PCR tests are so valuable: 02:00-03:10Changes in clinical detection rate and infectivity: 03:10-8:00Infectivity and exceptions to the rule: 08:00-09:00Sample collection methods, sensitivity and encouraging testing uptake 09:00-17:00The Broad Institute’s mass testing regimen: 17:00-21:00Developments during the pandemic and no RNA extraction PCR: 21:00-27:00Dreams for improving COVID-19 testing: 27:00- 30:00This episode was supported byRoche sequencing and life science. Follow these links to find out more about theLightCycler 480System andLightCycler 96 System.

Reproducibility in microbiomics

Season 2, Ep. 4
Returning to the contentious topic of the Reproducibility crisis –the inability of many study results to be replicated by different research groups or labs –this episode, supported by Zymo Research, zeros in on the topic within the field of microbiomics. Speaking to me about the topic is Raul Cano, Chief Scientific Officer at the BioCollective. Raul discusses the crisis and explains why it is prevalent in the emerging field of microbiomics before taking a look at the key role that microbiomics is beginning to play in the field of diagnostics, and how – if we make changes now – lasting improvements can be made in the field. Raul also lays out the three key areas that are holding back reproducibility in microbiomics and explains the actions that can be taken to improve the situation.Contents:Introduction: 00:00-02:00Explaining the reproducibility crisis: 02:00-02:55How microbiomics compare to other fields in terms of reproducibility: 02:55-04:05Challenges in reproducibility specific to microbiomics: 04:05-05:45The importance of reproducibility in microbiomics: 05:45-07:00Microbiomics in diagnostics: 07:00-10:00 Three key causes of poor reproducibility in microbiomics: 10:00-11:30Is detailed documentation improving in the life sciences? 11:30- 13:00Actions that can improve reproducibility: 13:00-14:15Fecal references: 14:15-16:05The realities of preparing fecal references: 16:05-18:1 5The reception to the new fecal references: 18:15-20:35References vs Standards: 20:35-22:15Standardizing microbiomics data: 22:15-23:20Assessing reproducibility in previous studies: 23:20-25:15Dreams for reproducibility in microbiomics: 25:15-26:55 This episode is supported by Zymo Research. If you would like more information about the fecal reference mentioned in the podcast, you can visit the product page here.

COG-UK: sequencing SARS-CoV-2 and detecting the novel variant B.1.1.7

Season 2, Ep. 2
This episode, supported by Tecan, takes a look at the role the COVID-19 Genomics UK Consortium (COG-UK) has played sequencing SARS-CoV-2 and surveying for COVID-19. To do this I speak to two key members of the consortium; Steve Paterson, Professor of genetics at the University of Liverpool and lead for the wastewater working group of COG-UK; and Josh Quick, Future Leaders Fellow at the University of Birmingham and lead at the COG-UK Sequencing working group.Steve discusses some of the techniques required to detect SARS-CoV-2 in wastewater, the challenges that such a vibrant sample can present, and gives his account of the part that wastewater surveillance played in the management of the new variant B.1.1.7.Josh provides us with further insight into the technologies used to sequence SARS-CoV-2, explains how he designed the ARTIC protocol for sequencing the virus and why it came to be so widely used. We go on to discuss the issues of limited lab consumables such as pipette tips and how you can make the most out of your limited lab supplies. Josh also describes the bizarre act of serendipity that aided in the discovery of the B.1.1.7 variant.ContentsIntroduction: 00:00-01:30Steve Paterson introduction: 01:30-02:15Introducing COG-UK: 02:15-03:20Becoming the wastewater working group lead: 03:20-04:35Key techniques and essential work in wastewater surveillance: 04:35-08:00Improving the sensitivity of sequencing and technological developments: 08:00-10:00Detecting new variants in wastewater screening: 10:00-12:15Learnings from the pandemic: 12:15-13:40Josh Quick introduction: 14:35-15:32The ARTIC protocol: 15:32-18:35Sequencing working group key techniques and key focuses 18:35-22:00Challenges of limited consumable supply and how to make the most of what you have got 22:00-24:30Learnings from the pandemic: 24:30-26:28New technologies, RC-PCR: 26:28-29:38Fantasy technology to assist with SARS-CoV-2 sequencing: 29:38-31:00Discovering the new variant B.1.17:31:00-33:30Conclusions: 33:30-34:40

The reproducibility crisis: where are we now and how can we progress?

Season 1, Ep. 11
In this episode I address the 'reproducibility crisis'. The concerning trend in the life sciences in which published papers and established principals are unable to be repeated by other researchers and, at times, even by the researchers who provided the initial results. To do this I speak to two researchers driving forward reproducibility in the life sciences, Marc Raphael from the United States Naval Research Laboratory, and Elizabeth Iorns, CEO of Science Exchange. Marc discusses his recent trial of independent verification and validation (IV&V) labs in a live science setting, highlighting the lessons learned from the experience and exploring the differences observed in attitudes and approaches to reproducibility between the life sciences and engineering, where mark began his career. Elizabeth discusses her work with Science Exchange and the reproducibility project: cancer biology. Elizabeth notes the key themes that have been exposed by the reproducibility project that are holding back the resolution of the crisis and explains what she thinks has worked so far in improving reproducibility. This leads to a fascinating discussion between Marc and Elizabeth, comparing and contrasting their experiences and noting the different challenges that they face in trying to improve the state of research in the life sciences. In fact, their camaraderie was so strong that interviewing these two was a real treat and for lots of it, it was much better for me to sit back and watch as they debated a topic that they are both clearly very passionate about!I hope you enjoy listening as much as I enjoyed recording this episode!*The Talking Techniques podcast is hosted and produced by Tristan Free for BioTechniques. We would love to here more from our listeners about your opinions on the podcast, what you would like to hear more of and what you think we could do better!Please get in contact on any of the platforms below.Twitter: @TristanFreefsgEmail: