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#29 — Ever wondered why you have to add protease inhibitors to your lysis buffer? In this episode of Mentors At Your Benchside, we’ll tell you all about protease inhibitors, why they’re important and how to use and store them. To learn more about protease inhibitors and to access the table with information about common protease inhibitors, read the full article on our website. [1] Read more about how protease inhibitors work, [2] learn about protein expression, [3] and find out what you need to consider during cell lysis. [4] Resources: 1. Protease Inhibitors 101: How They Work and How to Use Them. Available at: https://bitesizebio.com/58195/protease-inhibitors-101/ 2. How Proteases and Protease Inhibitors Work. Available at: https://bitesizebio.com/8845/how-proteases-and-protease-inhibitors-work/ 3. The Bitesize Bio Guide to Protein Expression – a Bitesize Bio eBook. Available at: https://bitesizebio.com/webinar/the-bitesize-bio-guide-to-protein-expression-a-bitesize-bio-ebook/ 4. Four Important Considerations for Your Cell Lysis. Available at: https://bitesizebio.com/40942/4-important-considerations-for-your-cell-lysis/

#28 — SDS-PAGE gels can be run at constant current, constant voltage, or constant power. But which is best? Listen to this episode of Mentors at Your Benchside to discover the differences between current, voltage, and power, and how they affect how your gels run. Visit the original article for a summary table of running your SDS-PAGE gels at a constant voltage, current, or power. [1] We've also got tips and tricks for casting the perfect SDS-PAGE gel, [2] or read a refresher on how SDS-PAGE works. [3] Resources: 1. Constant Current or Voltage in SDS-PAGE: The Great Debate. Available at: https://bitesizebio.com/51744/constant-current-or-voltage-in-sds-page/. 2. A Simple SDS-PAGE Gel Recipe and 10-Step Casting Protocol for Perfect Gels. Available at: https://bitesizebio.com/59429/sds-page-gel-recipe/. 3. How SDS-PAGE Works. Available at: https://bitesizebio.com/580/how-sds-page-works/.

#27 — Optimal conditions for DNA ligation reactions are a delicate balance between DNA molecules interacting and the enzymatic ligation reaction. In this episode, you'll discover our top 6 DNA ligation tips to improve the efficiency of your ligations and increase your cloning success rate! Read the full article to get more practical advice for optimizing DNA ligation reactions for cloning. [1] Want more cloning advice? Read about Cloning with Compatible Cohesive Ends, [2] and discover how to Remove Unwanted DNA from Vectors. [3] Also, find out why T4 DNA ligase is such a popular ligase and perhaps the only ligation enzyme you’ll ever need. [4] Resources: 1. DNA Ligation: How it Works & 6 Top Tips. Available at: https://bitesizebio.com/10279/how-dna-ligation-works/ 2. Assembling the Puzzle: Cloning with Compatible Cohesive Ends. Available at: https://bitesizebio.com/21716/assembling-the-puzzle-cloning-with-compatible-cohesive-ends/ 3. Tech Clinic #1: Removing Unwanted DNA from Vectors — beat Murphy’s Law. Available at: https://bitesizebio.com/10228/removing-unwanted-dna-from-vectors-how-to-beat-murphys-law/ 4. T4 DNA Ligase: The Only Ligase You’ll Ever Need? Available at: https://bitesizebio.com/46482/t4-dna-ligase-the-only-ligase-youll-ever-need/

#26 — Cryo-EM is a revolutionary imaging method that lets us see complex biostructures at higher and higher resolutions. But do you understand the mind-blowing science behind this technique? And what is cryo-electron microscopy, anyway? Why is the cryogenic aspect important, and how did it seemingly go from nothing to the big time? In the latest episode of Mentors At Your Benchside, we answer all of these questions and more! Check out the corresponding online article to access loads of follow-up resources to deepen your understanding of this topic.[1] Also, check out our related articles covering crucial sample preparation considerations for cryo-EM and its history from obscure to Nobel Prize winner. [2,3] Resources: 1. What Is Cryo-Electron Microscopy? Available at: https://bitesizebio.com/62871/what-is-cryo-electron-microscopy/ 2. Cryo-EM Sample Prep: 5 Crucial Considerations. Available at: https://bitesizebio.com/62619/cryo-em-sample-prep/ 3. A Short History of Cryo-Electron Microscopy: Available at: https://bitesizebio.com/62839/history-of-cryo-electron-microscopy/

#25 — Phenol extraction of DNA is a commonly used method for removing proteins from nucleic acids; for example, to remove proteins from cell lysate during genomic DNA preparation. It's commonly used, but not well understood. Listen to this episode to get a quick explanation of how phenol extraction of DNA works. Visit the original article to see handy diagrams on how phenol extraction works. [1] To get more help and advice on DNA extraction, read our related articles and discover how chloroform can clean up your phenol extractions, [2] learn how ethanol precipitation works, [3] and decide if ethanol or isopropanol is best for precipitating your samples. [4] Resources: 1. The Basics: How Phenol Extraction of DNA Works. Available at: https://bitesizebio.com/384/the-basics-how-phenol-extraction-works/ 2. Practical Application of Phenol-Chloroform Extraction. Available at: https://bitesizebio.com/3651/practical-application-of-phenol-chloroform-extraction/ 3. Ethanol Precipitation of DNA and RNA: How it Works. Available at: https://bitesizebio.com/253/the-basics-how-ethanol-precipitation-of-dna-and-rna-works/ 4. DNA Precipitation: Ethanol vs. Isopropanol. Available at: https://bitesizebio.com/2839/dna-precipitation-ethanol-vs-isopropanol/

#24 — How can you best prepare for a postdoc interview? Here we cover 10 questions that are often encountered during the job-hunting process. Preparing answers to these questions, alongside doing some background research about the research group and PI, can give you the edge in securing that postdoc. You can recap these top 10 questions anytime by visiting the full article on popular postdoc interview questions. [1] If you are looking for further interview advice, make sure you check out our top 10 interview tips, consider how you might concisely describe your research so far, and get advice on what to wear for the interview. Resources: 1. Postdoc Interview Preparation: Sample Questions and Answers. Available at: https://bitesizebio.com/10393/postdoc-interview-preparation-sample-questions-and-answers/ 2. 10 Great Tips To Make A Good Impression At Your Interview. Available at: https://bitesizebio.com/6696/10-great-tips-to-make-a-good-impression-at-your-interview/ 3. Can You Describe Your Research in 30 Seconds? 60? Available at: https://bitesizebio.com/13639/can-you-describe-your-research-in-30-seconds-60/ 4. GOT AN INTERVIEW! What to wear? What to wear? Available at: https://bitesizebio.com/6261/got-an-interview-what-to-wear-what-to-wear/

#23 — Creativity in Science: How a Good Imagination Can Help Your Research If you're stuck in a research rut in the lab, developing your creativity can help you find new solutions to current problems that may be impeding your scientific progress. In this episode, we explore 3 important questions you should consider to help spark your creativity in the lab and provide 8 simple tips to help achieve this. Read the full article on boosting your creativity in science. [1] If you are looking for further advice, make sure you listen to our Happy Scientist Podcast episode on Imagination, which covers even more ways to boost your imagination and creativity. [2] Consider the importance of allowing yourself time to think and reflect. [3] Finally, listen to this episode of the Microscopists featuring Eric Betzig, where the Nobel Laureate reveals how his biggest ideas came during his two periods of unemployment. [3] Resources: 1. Creativity in Science: How a Good Imagination Can Help Your Research. Available at: https://bitesizebio.com/63742/creativity-in-science/ 2. Episode 33 — How to Foster Imagination. Available at : https://bitesizebio.com/podcast/episode-33-how-to-foster-imagination/ 3. Time to Think. Available at: https://bitesizebio.com/146/time-to-think/ 4. Eric Betzig (University of California). Available at: https://bitesizebio.com/podcast/eric-betzig/

#22 — Research often requires you to use dangerous chemicals. From caustic acids and bases to pH solutions and toxic reducing agents, chemical hazards abound in the lab. In this episode, we'll take a look at ten dangerous chemicals in the lab and the harm they can cause you. Want to develop your understanding of chemical safety? Why not read the full article, [1] where you'll find lots of helpful resources and further reading? If you fancy a deeper dive into all aspects of lab safety, then check out the Bitesize Bio lab safety eBook. [2] If you're a bit rusty on your good laboratory practice, definitely read 10 Stupid Lab Safety Mistakes [3] and see how many sins you commit. And finally, we all rely on our PPE for protection and safety. Be sure to take a look at our 10 Common PPE Sins for a quick brush-up. [4] Resources: 1. Ten Bad Chemicals in the Lab and What They do to You! Available at: https://bitesizebio.com/10470/ten-bad-chemicals-in-the-lab-and-what-they-do-to-you/ 2. The Bitesize Bio Guide to Lab Safety eBook. Available at: https://bitesizebio.com/wp-content/uploads/2021/08/The-Bitesize-Bio-Guide-to-Lab-Safety.pdf 3. 10 Stupid Lab Safety Mistakes. Available at: https://bitesizebio.com/1899/stupid-lab-safety-mistakes/ 4. 10 Common PPE Sins. Available at: https://bitesizebio.com/9850/10-common-ppe-sins/

#21 — Do you know the difference between cell confluency and cell number? Can you measure cell confluency accurately? Cell confluency measurements are essential in cell-based experiments. Listen to this episode to learn what cell confluency is and the different quantification methods to measure it. Read the original article to get visual guides to help you understand and measure your cell confluency accurately. [1] Do you want more information to help you perfect your cell culture? Check out related articles to learn how to get the right passage number for your cells and how to culture adherent cells like HEK293s. [2,3] Resources: 1. How Confluent Are Your Cells? A Beginner’s Guide to Measuring Cell Culture Confluency. Available at: https://bitesizebio.com/63887/cell-confluency/ 2. What’s in a Number: Getting the Right Passage in Cell Culture. Available at: https://bitesizebio.com/13685/cell-culture-passage-number-explained/ 3. What the HEK? A Beginner’s Guide to HEK293 Cells. Available at: https://bitesizebio.com/45489/guide-to-hek293-cells/

#19 — Listen to this episode to discover what alternative splicing is, the history of how it was discovered, and get key examples of splicing in action. Read the full article to learn more about alternative splicing, including a visual diagram of how splicing works. [1] Read our related articles to find out how alternative splicing can affect your experiments and discover methods for detecting splice variants. [2,3] Resources: 1. What is Alternative Splicing, and Why Is It Important? Available at: https://bitesizebio.com/10148/what-is-alternative-splicing-and-why-is-it-important/ 2. How Can A Single Mutation Affect Splicing Regulation? Available at: https://bitesizebio.com/10307/how-can-a-single-mutation-affect-splicing-regulation/ 3. How to Detect Alternative Splicing Variants. Available at: https://bitesizebio.com/10138/how-to-detect-alternative-splicing-variants/

#18 — Why are SDS-PAGE gels run vertically? What would happen if we ran them horizontally? Has anyone ever tried? Discover the answers to these questions and more in this episode of Mentors At Your Benchside. To see what others are saying and join in the conversation, head over to the comments section in the original article on why SDS-PAGE is run vertically. [1] If this episode has ignited your desire for answers, Check out some of our related articles to learn the how SDS-PAGE works and discover whether running your SDS-PAGE gels with constant current or constant voltage is best. [2,3] Resources: 1. Why Is SDS-PAGE Run Vertically? Here are 3 Great Answers. Available at: https://bitesizebio.com/10699/why-is-sds-page-run-vertically/ 2. How SDS-PAGE Works. Available at: https://bitesizebio.com/580/how-sds-page-works/ 3. Constant Current or Voltage in SDS-PAGE: The Great Debate. Available at: https://bitesizebio.com/51744/constant-current-or-voltage-in-sds-page/

#17 — Why do some plasmids give a better protein yield than others? It may have a lot to do with the plasmid copy number. In this episode, we talk about what plasmid copy number means, why it is important and how you can manipulate it to get the most out of your experiments. To learn more about plasmid copy number, read the full article on our site. [1] Resources: 1. https://bitesizebio.com/22824/how-to-manipulate-plasmid-copy-number/

#16 — Is your viva on the horizon? Many people think viva exams are going to be painful. But what if you could make your viva go smoothly and maybe even enjoy it? Listen to our top 10 viva tips to get advice from a viva survivor. To learn more about surviving your viva, read the full article on our site. [1] Resources: 1. https://bitesizebio.com/10109/top-10-tips-for-viva-success/

#15 — In the last few years, CRISPR-Cas9 has become a popular genome editing tool, but have you ever wondered where it came from? In this episode, we explore the history of CRISPR-Cas9, from the first CRISPR discovery in 1987 to current applications in targeted genome and epigenome editing. Read the full article to learn more about the history of CRISPR-Cas9 and to see a timeline of discoveries. [1] Resources: 1. https://bitesizebio.com/47927/history-crispr/

#14 — Do you know how accurate and precise your measurements are? Imprecise and inaccurate measurements can have a dramatic impact on your results. Listen to get 8 top tips for making your measurements more accurate and precise. To learn more about accuracy and precision, read the full article on our site. [1] Resources: 1. https://bitesizebio.com/55470/accuracy-and-precision/

#13 — There are now hundreds of different tailored fluorescent proteins to choose from that can improve your experiments in the lab. What are the key considerations when choosing a fluorescent protein for your experiment? Should you consider using a fancy new fluorescent protein? Where can you find the best resources to help you out? Find out the answers to these and more in this episode of Mentors At Your Benchside. Please read the full article, which includes our super helpful table of fluorescent proteins and their properties. [1] Resources: 1. https://bitesizebio.com/54287/choosing-a-fluorescent-protein/

#12 — As a follow-up to our episode about ethanol precipitation of DNA and RNA, [1] this episode explains the differences between DNA precipitation in ethanol and isopropanol, helping you to figure out which method is the best choice for your experiment. Read the full article for handy protocol tips, the differences between using ethanol and isopropanol, and when to use each method. [2] Resources: 1. https://bitesizebio.com/2007/12/04/the-basics-how-ethanol-precipitation-of-dna-and-rna-works/ 2. https://bitesizebio.com/2839/dna-precipitation-ethanol-vs-isopropanol/

#11 — Fluorescence is undoubtedly one of the most important and useful tools in a biologist’s toolbox. But do you actually know how fluorescence works? In this episode, discover what the three steps of fluorescence are, and how fluorescence can be used in flow cytometry. Read the full article for a breakdown of the key points of fluorescence. [1] Resources: 1. https://bitesizebio.com/32973/fluorescent-molecules/

#10 — As you probably know, there are different types of ethanol found in biology labs. It's a versatile solvent used in dozens of experiments and procedures, including disinfection, DNA precipitation, and tissue dehydration. But which type of ethanol should you use for your application? What's the difference between the types of ethanol in your lab? And how do you handle it appropriately? In this episode, we answer these questions, providing you with the information you need to keep your research producing top-notch results. Read the full article on Which Type of Ethanol You Should Use for links to additional resources on laboratory applications of ethanol. [1] Resources: 1. https://bitesizebio.com/13518/which-type-of-ethanol-should-i-use/

#9 — One of the most commonly used cell lines in molecular biology labs is the Human Embryonic Kidney HEK293 cell line. Listen to this episode to find out all about the history of HEK293 cells and how to work with them. Read the full article for more details about working with HEK293 cells. [1] Resources: 1. https://bitesizebio.com/45489/guide-to-hek293-cells/