Mentors at Your Benchside

#79 — Are you struggling to keep your proteins "happy" and active for your experiments? In this episode of Mentors At Your Benchside, we dive into the five critical elements you need to design the ideal protein purification buffer: pH, the buffer system, salt concentration, reducing agents, and stabilizing additives. [1] Learn how buffers work, [2] and how to navigate the delicate balance of these factors to prevent protein aggregation and keep your purified proteins soluble and active. [3] Tune in for a simplified guide to mastering your protein purification buffer. Resources: 1. 5 Ingredients for the Perfect Protein Purification Buffer. Available at: https://bitesizebio.com/7893/how-to-design-the-perfect-protein-purification-buffer/ 2. How Do Buffers Work? Available at: https://bitesizebio.com/8478/how-do-buffers-work/ 3. The Definitive Guide to pH, pKa, and pI. Available at: https://bitesizebio.com/9425/what-is-ph-pka-pi/

#78 — Want to visualize if your proteins interact in live cells? FRET is the answer. In this Mentors at Your Benchside episode, we explain how FRET works, why it's great for studying protein–protein interactions, and why it is not actually named Fluorescence Resonance Energy Transfer. Visit the original article to see helpful diagrams, [1] brush up on how fluorescent molecules work, [2] and read up on how to measure FRET for the different methods and practical tips for choosing FRET pairs. [3] Resources: 1. How FRET Works: A Guide to Visualizing Protein Interactions. Available at: https://bitesizebio.com/23012/how-fret-works/ 2. How Fluorescent Molecules Work. Available at: https://bitesizebio.com/32973/fluorescent-molecules/ 3. How to Measure FRET. Available at: https://bitesizebio.com/23295/how-to-measure-fret/

#77 — Have you ever thought about accessibility in science? We don’t always present our science in ways that are accessible to everyone. Nor is lab-based science always accessible. In this episode of Mentors At Your Benchside, we explore what accessibility is and highlight how we can all make science more accessible and inclusive. Visit the original article for helpful resources and to revisit the key definitions. [1] Make your presentations more accessible using the color blindness simulator, [2] the accessible color pallets, [3] and the list of accessible fonts. [4] Resources: 1. Accessibility in Science: How Can We Make Science More Accessible? Available at: https://bitesizebio.com/65863/accessibility-in-science/ 2. Coblis —Color Blindness Simulator. Available at: https://www.color-blindness.com/coblis-color-blindness-simulator/ 3. Coloring for Colorblindness: Color Palette. Available at: https://davidmathlogic.com/colorblind/#%23D81B60-%231E88E5-%23FFC107-%23004D40 4. My favorite free fonts for print disabilities. Available at: https://www.perkins.org/resource/my-eight-favorite-free-fonts-print-disabilities/

#76 — Do you know how the first cells were identified? Or who discovered them? What about why they are called cells? Discover the fascinating history in this enlightening Mentors At Your Benchside episode. Visit the original article for a timeline of cell biology, [1] discover the most commonly used cell lines, [2] and find out why HeLa cells are surrounded by controversy. [3] Resources: 1. History of Cell Biology. Available at: https://bitesizebio.com/166/history-of-cell-biology/ 2. Top 5 of the Most Commonly Used Cell Lines. Available at: https://bitesizebio.com/33473/top-5-of-the-most-commonly-used-cell-lines/ 3. The Story Behind Your Cell Culture. Available at: https://bitesizebio.com/7647/the-story-behind-your-cell-culture/

#75 — Counting your cultured cells is vital to seeding the right density for your experiments, harvesting an appropriate amount of downstream experiments, preparing cells for flow cytometry, and more. Luckily it's pretty easy with a hemocytometer. In this episode of Mentors At Your Benchside, we talk you through the four steps of counting cells using a hemocytometer, including best practices so your count is accurate. Visit the original article for helpful images of the hemocytometer grid, [1] read up on why cell counting is so important, [2] and get your free cell culture posters, including a handy hemocytometer poster. [3] Resources: 1. Cell Counting with a Hemocytometer: As easy as 1, 2, 3… Available at: https://bitesizebio.com/13687/cell-counting-with-a-hemocytometer-easy-as-1-2-3/ 2. A Numbers Game: the ‘How’ and ‘Why’ of Counting Cells. Available at: https://bitesizebio.com/22758/a-numbers-game-the-how-and-why-of-counting-your-cells/ 3. Bitesize Bio's Handy Cell Culture Posters. Available at: https://bitesizebio.com/cell-culture-posters/

#74 — Understanding how a technique works make it simpler to troubleshoot when things go wrong in your experiments. Learn how alkaline lysis works in this short and simple step-by-step run-through of the process. Check out the original article for links to helpful resources, [1] discover five ways to clean up a DNA sample, [2] and get tips on preparing your vectors for gene cloning. [3] Resources: 1. Lab Basics: How The Alkaline Lysis Method Works. Available at: https://bitesizebio.com/180/the-basics-how-alkaline-lysis-works/ 2. 5 Ways to Clean Up A DNA Sample. Available at: https://bitesizebio.com/142/5-ways-to-clean-up-a-dna-sample/ 3. 5 Tips on Vector Preparation for Gene Cloning. Available at: https://bitesizebio.com/13500/cloning-tips-vector-prep/

#73 — PCR is a fundamental technique all biologists rely on, and, for qPCR, we can construct a standard curve that tells us how good or bad our primers are. In this episode, learn all about qPCR standard curves, what they tell you about your primers, and what to try if something doesn't look correct on your standard curve. Check out the corresponding online article for an example qPCR curve. [1] For more PCR essentials, download our free eBook that explains everything you need to know. [2] And to drill specifically into the efficiency of your PCR, read our article on determining qPCR efficiency. [3] Resources: 1. The qPCR Standard Curve: The Key to Good qPCR Data. Available at: https://bitesizebio.com/31470/obligate-qpcr-standard-curve/ 2. The Fundamentals of qPCR and RT-qPCR. Available at: https://bitesizebio.com/the-fundamentals-of-qpcr-and-rt-qpcr/ 3. Important Considerations for Determining qPCR Efficiency. Available at: https://bitesizebio.com/3177/determining-qpcr-efficiency/

#72 — Research requires imagination and strategy, and helpful distractions can give us the mental rest we need to recharge. Fun hobbies for scientists can provide inspiration, creativity, fitness, articulacy, and a much-needed break. Discover our top 10 hobbies for scientists, and find a new hobby that could also benefit your research. Visit the original article for links to related resources [1] and discover why creativity is so important in science. [2] For more helpful tips to boost your research, read our article on how to give an engaging talk [3] or take our course on critical thinking. [4] Resources: 1. 10 Fun Hobbies for Scientists. Available at: https://bitesizebio.com/74251/hobbies-for-scientists/ 2. Creativity in Science: How a Good Imagination Can Help Your Research. Available at: https://bitesizebio.com/63742/creativity-in-science/ 3. How to Give a Great Scientific Talk and Engage Your Audience. Available at: https://bitesizebio.com/29880/great-scientific-talk/ 4. How to Critically Review a Scientific Manuscript. Available at: https://bitesizebio.com/courses/critical-review/

#71 — A good histology slide can give you beautiful, revealing microscope images of your precious tissue samples. But what goes into preparing slides for histology? Whether you're new to the game, have only ever sent your samples off for slide preparation, or need a refresher, this episode explains how histology slides are prepared. Check out the corresponding article for links to related histology resources. [1] Plus, learn about the different fixes for histology [2] and the various ways you can section tissues. [3] Plus, check out our histology hub for more histology content, from free posters and guides to articles and a jargon-busting glossary of the common terms. [4] Resources: 1. How Histology Slides Are Prepared. Available at: https://bitesizebio.com/13398/how-histology-slides-are-prepared/ 2. 4 Fixatives for Histology and Cytometry. Perfect Your Preservation. Available at: https://bitesizebio.com/22141/fixation-and-flow-cytometry/ 3. The Perfect Slice: Preparing Tissue Samples For IHC. Available at: https://bitesizebio.com/13396/the-perfect-slice-preparing-tissue-samples-for-ihc-2/ 4. Histology Made Simple: An Easy Guide for Bioscientists. Available at: https://bitesizebio.com/histology/

#70 — Sterilization is a critical technique in the biology lab. It keeps your cell lines free from contamination, allows safe disposal of used items, and prevents breakouts of phage! In this episode, we discuss six sterilization techniques and explain how they work. Read the original article for additional resources on basic lab techniques. [1] If you're curious about how UV light damages DNA, read this article. [2] And check this article out for the lowdown on filtration. [3] Resources: 1. 6 Laboratory Sterilization Methods. Available at: https://bitesizebio.com/853/5-laboratory-sterilisation-methods/ 2. How UV Light Damages DNA and the Havoc it Can Cause to Your Experiments. Available at: https://bitesizebio.com/36762/how-uv-light-damages-dna/ 3. How Filtration Works: A Short Guide for Biologists. Available at: https://bitesizebio.com/59280/how-filtration-works/

#69 — Figures are a fundamental way to communicate science and are essential components of journal articles. In this episode, learn the differences between vector and raster image types, hear the pros and cons of various common file types, and get the lowdown on common parameters such as color models and DPI. Check out the original article for an easy table that summarizes the key differences between image types. [1] Plus, if you want to make beautiful images of proteins and DNA, read our step-by-step tutorial, [2] and if you need some inspiration, check out our article on the science of figure creation for ideas to make your figures stand out. [3] Resources: 1. The 2 Types of Digital Images: Tools to Prepare Stunning Images for Publication. https://bitesizebio.com/43785/an-introduction-to-digital-images-in-publications/ 2. Learn to Draw a Molecule in PyMOL™ in 8 Easy Steps. Available at: https://bitesizebio.com/54445/molecular-visualization-tool/ 3. The Art and Science of Figure Creation: Think BIG to see Small. Available at: https://bitesizebio.com/30920/art-science-figure-creation/

#68 — Controls are fundamental to getting meaningful data. Especially in long, drawn-out experiments like immunofluorescence imaging. In this episode, we explain 5 types of immunofluorescence control, what they tell you, and why they are critical for your experiments. Check out the original article for links to loads of related resources [1], and you can access all this information as a beautiful poster for your lab. [2] Resources: 1. 5 Controls for Immunofluorescence: A Beginner’s Guide. Available at: https://bitesizebio.com/44370/controls-for-immunofluorescence-a-beginners-guide/ 2. Bitesize Bio's Ultimate Immunofluorescence Troubleshooting Guide. Available at: https://bitesizebio.com/immunofluorescence-troubleshooting-guide/

#67 — What are those mysterious extra water molecules at the end of some chemical names? Do they matter to your experiments? And what should you do when they aren't water but are something more intrusive like HCl? In this episode, we answer all those questions and tell you everything you need to know about water of crystallization. Check out the original article for handy figures that illustrate water of crystallization in real chemicals. [1] And if you want more info on the chemistry that happens in biology labs, check out our article explaining what common buffer additives do [2] and read this article to learn more about pH, pI, and pKa. [3] Resources: 1. What is Water of Crystallization? Everything You Need to Know. Available at: https://bitesizebio.com/73932/water-of-crystallization/ 2. Just What Do All These Additives Do? Available at: https://bitesizebio.com/19420/just-what-do-all-these-additives-do/ 3. The Definitive Guide to pH, pKa, and pI. Available at: https://bitesizebio.com/9425/what-is-ph-pka-pi/

#66 — Are your plasmid yields low? Not sure what is considered a good yield? Need help boosting the amount of plasmid you get from your preps? In this episode, we discuss what is a good and bad yield, why your plasmid preps might be sub-optimal, and how you get better yields. Read the original article for an easy protocol for growing unsaturated cultures. [1] Discover the differences between DNA precipitation and ethanol vs. isopropanol in our article [2] and watch Eppendorf's webinar on centrifugation basics. [3] Resources: 1. Better Plasmid Purification: 11 Reasons Your Plasmid Yield is Low. Available at: https://bitesizebio.com/13514/boost-plasmid-yield/ 2. DNA Precipitation: Ethanol vs. Isopropanol. Available at: https://bitesizebio.com/2839/dna-precipitation-ethanol-vs-isopropanol/ 3. Essentials in Centrifugation - Better Safe than Sorry! Available at: https://events.bitesizebio.com/essentials-in-centrifugation-1/

#65 — Delegation is not just for managers! No matter where you are in your research career, there is an opportunity to delegate. In this episode, we discuss how you can simplify your lab life by delegating, who you can delegate to, and how to do it effectively and without offending anyone! Read the full article to review everything covered in this episode. [1] If you want a deeper dive into how and when to delegate, listen to the When and How to Delegate episode of The Happy Scientist podcast. [2] And if you're a manager looking to delegate down, read our top 10 tips for being a good boss. [3] Resources: 1. The Importance of Delegation for Scientists. Available at: https://bitesizebio.com/61711/delegation-for-scientists/ 2. When and How to Delegate. Available at: https://thehappyscientist.bitesizebio.com/episodes/ep-35-when-and-how-to-delegate 3. 10 Ways to Be a Good Boss. Available at: https://bitesizebio.com/1869/10-ways-to-be-a-good-boss/

#64 — Like many experiments, doing a Western blot typically means comparing half a dozen online protocols, failing completely, triple-checking the recipes of your more suspicious buffers, then remaking them and starting again anyway. To ease the pain, this episode gives you a reliable ECL reagent recipe so you can prepare your own fresh (and private) stash anytime you need. We also explain what all the components do. Check out the corresponding article to see a mechanism for the oxidation of luminol and links to papers discussing alternative enhancers. [1] If you want better Western blot transfers, read our top three tips. [2] And better yet, we've put this ECL reagent recipe, an adaptable Western blot protocol complete with buffer recipes, and loads of troubleshooting tips together in a cheat sheet to stick in your lab book. Download it for free here. [3] Resources: 1. How To Make ECL Reagent: 4 Ingredients, Better Blots. Available at: https://bitesizebio.com/8970/how-to-make-your-own-ecl/ 2. 3 Hot Tips to Optimize Your Western Blot Transfers. Available at: https://bitesizebio.com/7356/optimize-your-western-blot/ 3. Bitesize Bio's Western Blot Protocol Cheat Sheet. Available at: https://bitesizebio.com/western-blot-protocol/

#63 — Are you fed up with Western blots that look like your pen leaked in your pocket? We all are. That's why we've put together our three favorite tips for better Western blot transfers. Visit the full article for a handy figure illustrating the elements of the transfer stack. [1] You can also learn more about the different approaches to blotting. [2] Plus, we've got an easy ECL reagent recipe for you too. [3] Resources: 1. 3 Hot Tips to Optimize Your Western Blot Transfers. Available at: https://bitesizebio.com/7356/optimize-your-western-blot/ 2. 3 Approaches to Western blot Transfer: Available at: https://bitesizebio.com/8274/3-approaches-to-western-blot-transfer/ 3. How To Make ECL Reagent: 4 Ingredients, Better Blots. Available at: https://bitesizebio.com/8970/how-to-make-your-own-ecl/

#62 — Do you want to use a cell line but are unsure where to start? Or perhaps you’re just curious about the most commonly used cell lines. Listen to our top 5 most commonly used cell lines to get a feel for the cells that many researchers turn to. Read the original article, [1] brush up on the harrowing origins of the HeLa cell line, [2] and get advice on how to pick the perfect cell line for your research. [3] Resources: 1. Top 5 of the Most Commonly Used Cell Lines. Available at: https://bitesizebio.com/33473/top-5-of-the-most-commonly-used-cell-lines/ 2. The Story Behind Your Cell Culture. Available at: https://bitesizebio.com/7647/the-story-behind-your-cell-culture/ 3. The Do’s and Dont’s of Choosing a Cell Line. Available at: https://bitesizebio.com/46957/choosing-a-cell-line/

#61 — Fed up with fighting to put on another pair of gloves with sticky hands? Faced the embarrassment of giving a sweaty handshake? You need to listen to our top tips for handling sweaty glove hands in the lab. Visit the full article to leave your comments on handling sweaty glove hands, [1] and check out our guide to choosing the right glove. [2] Resources: 1. Get a Grip: Dealing with Sweaty Glove Hands. Available at: https://bitesizebio.com/44798/get-a-grip-dealing-with-sweaty-glove-hands/ 2. Guard Yourself with Our Great Lab Glove Selection Guide. Available at: https://bitesizebio.com/22956/guard-yourself-with-our-guide-to-gloves/

#60 — There's nothing quite as frustrating as imaging a western blot only to find your bands are a wonky mess, your blot is covered in bubbles, or your bands are missing entirely. We've got a solution to help: Ponceau S. This stain gives a midpoint step that can help narrow down the cause of your problems. Listen to this episode of Mentors At Your Benchside to discover a step-by-step guide for performing Ponceau S staining and how to interpret what the staining means when troubleshooting your failed blots. Visit the full article to get a written protocol and see examples of Ponceau S staining. [1] Plus, for more help and advice on performing western blots, download our Free Western Blot Protocol Cheat Sheet. [2] Resources: 1. What Ponceau S Staining Can Reveal About Your Western Blot. Available at: https://bitesizebio.com/72803/ponceau-s-staining/ 2. Bitesize Bio's Western Blot Protocol Cheat Sheet. Available at: https://bitesizebio.com/western-blot-protocol/