#112 — Isoelectric focusing (IEF) is a powerful technique distinct from the more familiar SDS-PAGE, [1,2] tailored for separating proteins or peptides based on their isoelectric points (pI). [3]

This method capitalizes on the migration of charged molecules through a stable pH gradient until they reach a zone where their net charge is zero, halting their movement.

Essential to setting up an IEF experiment are the IPG strips, equipped with a pH-responsive gel, and the sample, often mixed with carrier ampholytes for efficient migration.

IEF's utility spans various applications, from enhancing 2D-PAGE protein separations to analyzing post-translational modifications and preparing samples for mass spectrometry. [4]

Discover more about how this technique can advance your research and streamline your experimental workflows.

Resources:
1. How SDS-PAGE Works. Available at: https://bitesizebio.com/580/how-sds-page-works/
2. Isoelectric Focusing: A Simple Way to Enhance Your Protein Separation. Available at: https://bitesizebio.com/26811/isoelectric-focusing-separation-proteins-peptides/
3. The Definitive Guide to pH, pKa, and pI. Available at: https://bitesizebio.com/9425/what-is-ph-pka-pi/
4. Detecting Post-Translational Modifications. Available at: https://bitesizebio.com/49322/detecting-post-translational-modifications/

#111 — In this episode, we dive deep into the fascinating world of microbes and their revolutionary applications in biotechnology. From environmental solutions to breakthroughs in health and medicine, microbes hold the key to some of the most advanced scientific developments.

Discover how these microscopic organisms are transforming industries and pushing the boundaries of what's possible in science and technology. We’ll explore the latest research and applications that are shaping the future.

For an in-depth look at this topic, make sure to read the corresponding article on our website. [1] Dive into an application you almost certainly know about, yeast two-hybrid assays, [2] and check this article out to learn how to increase protein expression in plants. [3]

Resources:
1. Top 10 Uses of Microbes in Biotechnology. Available at: https://bitesizebio.com/48446/microbes-biotechnology/
2. Split Ubiquitin Yeast Two-Hybrid. Available at: https://bitesizebio.com/25286/split-ubiquitin-yeast-two-hybrid/
3. P19 to the Rescue: How to Increase Protein Expression in Agroinfiltration. Available at: https://bitesizebio.com/29964/p19-increase-expression-agroinfiltration/

#102 — Fluorescence microscopy images not only look great but also allow us to get a better understanding of cells, structures, and tissues. And confocal laser scanning microscopy lets us construct 3D images from 2D micrographs.

In this episode, learn the basic principles of confocal laser scanning microscopy, how the microscopes work, and some of its applications in bioscience and beyond.

Check out the corresponding article for a handy confocal laser scanning microscope diagram. [1] Learn about the Airy unit [2] and check out our guide to choosing a fluorescent protein for your experiments. [3]

Resources:
1. Confocal Laser Scanning Microscopy Explained In 3 Easy Steps. Available at: https://bitesizebio.com/19958/confocal-laser-scanning-microscopy/
2. Rubbing Your Microscope’s Eyes: A Guide to Optical Resolution. Available at: https://bitesizebio.com/13415/rubbing-your-microscopes-eyes-a-guide-to-optical-resolution/
3. The Ultimate Guide to Choosing a Fluorescent Protein. Available at: 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 [2] for handy protocol tips, the differences between using ethanol and isopropanol, and when to use each method.

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 [1] for a breakdown of the key points of fluorescence.

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 [1] for links to additional resources on laboratory applications of ethanol.

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 [1] for more details about working with HEK293 cells

Resources:
1 - https://bitesizebio.com/45489/guide-to-hek293-cells/

#8 — You probably have or will use SDS-PAGE at some point to separate proteins, but do you really understand how this technique works? Knowing how SDS-PAGE works means you can tweak and troubleshoot your technique as well as impress your supervisor and lab mates.

In this episode, we take you through how SDS-PAGE works, including what SDS does, why you need to use a reducing agent like DTT or beta-mercaptoethanol, and the critical importance of the stacking gel.

Read the full How SDS-PAGE Works article [1] to see helpful visuals for how this technique works and access the table showing the protein sizes that different acrylamide percentages can separate.

Expand your knowledge by buffing up on laemmli buffer [2] and get our Guide to Gradient Gels. [3] If you pour your own SDS-PAGE gels, take a deep dive into the chemistry of how gels work and learn how to pour perfect gels every time with our  Simple SDS-PAGE Gel Recipe with 10-Step Casting Protocol. [4]

Resources:
1 - https://bitesizebio.com/580/how-sds-page-works/
2 - https://bitesizebio.com/44540/laemmli-buffer-what-is-it-for-anyway/
3 - https://bitesizebio.com/47184/gradient-gels/
4 - https://bitesizebio.com/59429/sds-page-gel-recipe/

#7 — In this episode, we cover how jellyfish were able to have a huge impact on biology through the discovery and development of GFP.

Using GFP is now ubiquitous in pretty much all fields in biology, and we'll take you through how three Nobel Laureates developed this valuable research tool. [1]

Many of the applications for GFP are microscopy-based, and we'll discuss how you can utilize GFP for translational and transcriptional fusions, FRAP, FLIP and FRET experiments in the lab. [2,3]

Read the full article for more useful links, hints, and tips on using GFP in your experiments. [4]

Resources:
1. Press release. NobelPrize.org. Nobel Prize Outreach AB 2022. Fri. 1 Jul 2022. Available at: https://www.nobelprize.org/prizes/chemistry/2008/press-release/
2. Fun With FRAP! Fluorescence Recovery After Photobleaching for Confocal Microscopy. Available at: https://bitesizebio.com/19946/fun-with-frap-fluorescence-recovery-after-photobleaching-for-confocal-microscopy/
3. Using Flow Cytometry for Fluorescence Resonance Energy Transfer. Available at: https://bitesizebio.com/21935/using-flow-cytometry-for-fluorescence-resonance-energy-transfer/
4. How a Jellyfish Changed Biology: the Discovery and Development of GFP. Available at: https://bitesizebio.com/13390/gfp/

#6 — Ethanol precipitation is a commonly used technique for concentrating and de-salting nucleic acid preparations in an aqueous solution.

In this episode, we'll bring you up to speed on how ethanol precipitation works, including the importance of solubility, the roles of salt, ethanol, and temperature, plus a few helpful tips on the side.

Read the full article [1] to learn more about the ins and outs of ethanol precipitation and other DNA clean-up approaches.

Resources:
1. https://bitesizebio.com/253/the-basics-how-ethanol-precipitation-of-dna-and-rna-works/

#5 — Have you ever chilled beer on dry ice from the lab? Does your wardrobe consist mainly of free t-shirts from companies at conferences [1]? Have you ever wondered what the distilled water in the lab tastes like [2]?

If any of these questions apply to you, then tune in to this episode, as we cover more funny and telling signs that you're a true scientist.

Read the full article [3] to discover the difference between a Science Fan and a bona fide Scientist

Resources:
1. https://bitesizebio.com/31887/scientific-conferences-how-where-to-go/
2. https://bitesizebio.com/33270/types-of-water-lab/
3. https://bitesizebio.com/36753/20-telling-signs-scientist/

#4 — PCR is a useful lab technique used to copy, sequence or quantify DNA. If you're new to PCR, this episode is for you. We cover the five things you need to get started with PCR and explain how to run a polymerase chain reaction.

Read the full article to learn more about setting up a PCR. [1]

Learn more about Kary Mullis and the invention of PCR, and about the first thermocycler "Mr Cycle". [2, 3, 4] For practical tips, see our article on the top tips for primer design. [5]

Resources:
1. What is PCR? The Beginner’s Guide. Available at: https://bitesizebio.com/19132/pcr-basics-what-is-pcr/
2. Kary B. Mullis – Facts. NobelPrize.org. Nobel Prize Outreach AB 2022. Fri. 1 Jul 2022. Available at: https://www.nobelprize.org/prizes/chemistry/1993/mullis/facts/
3. The Invention of PCR. Available at: https://bitesizebio.com/13505/the-invention-of-pcr/
4. Mr. Cycle, Thermal Cycler. Available at: https://americanhistory.si.edu/collections/search/object/nmah_1000862
5. How to Survive a Difficult PCR. Available at: https://bitesizebio.com/37205/how-to-survive-a-difficult-pcr/

#3 — In this episode of Mentors At Your Benchside, listen to a short history of cryo-electron microscopy—the Nobel Prize-winning technique revolutionizing molecular and structural biology.

The journey takes us from the inception of cryo-EM- obscure, inferior, and derided- to the present-day competition for access to the incredibly powerful Krios microscopes.

It's all about the power of an image.

And speaking of images, be sure to read A Short History of Cryo-Electron Microscopy for a graphical timeline and some stunning cryo-EM structures. [1]

Fascinating, cross-disciplinary science underpins cryo-EM—the technique is as broad as it is popular. To expand your understanding of the fundamental science behind it, Read What Is Cryo-Electron Microscopy? A Brief Introduction. [2] For tips and tricks on getting your sample ready for a cryo-EM experiment, check out Cryo-EM Sample Prep: 5 Crucial Considerations. [3] And for a simple illustration of the stunning structures cryo-EM produces, browse the Electron Microscopy Data Bank. [4]

Resources:
1. A Short History of Cryo-Electron Microscopy. Available at: https://bitesizebio.com/62839/history-of-cryo-electron-microscopy/
2. What Is Cryo-Electron Microscopy? A Brief Introduction. Available at: https://bitesizebio.com/62871/what-is-cryo-electron-microscopy/
3. Cryo-EM Sample Prep: 5 Crucial Considerations, Available at: https://bitesizebio.com/62619/cryo-em-sample-prep/
4. Electron Microscopy Data Bank. Available at: https://www.ebi.ac.uk/emdb/

#101 —  Discover what it takes to become an expert at getting funded, from simple habits such as summarizing what you read in the literature, to big steps such as organizing your very own conference to establish your name in your field.

With over 30 years of experience as a biomedical engineering researcher seeking grants, Joel Berry, Founder, and Chief Scientist at Astound Research, shares his hard-won insights on strategizing your approach to seeking grants. [1] Take a listen to what he has to say.

If you're struggling to keep up to date with the literature in your field, read our tips for staying on top of new publications [2] and get more grant writing advice from a grant reviewer! [3]

Resources:
1. How to Become an Expert at Getting Funded. Available at: https://bitesizebio.com/77308/how-to-become-an-expert-at-getting-funded/
2. Useful Tips to Keep on Top of New Literature. Available at: https://bitesizebio.com/10532/search-engines-for-literature-searching/
3. What I Learned as a Grant Reviewer. Available at: https://bitesizebio.com/30909/learned-grant-reviewer/

#2 — Micropipettes are the bread and butter instrument of a lab, helping you to accurately dispense minute amounts of liquid. But these tools are only trustworthy if they are well maintained and handled appropriately. Listen to the latest episode of Mentors At Your Benchside to uncover top tips for precise pipetting.

Want to know more about correct pipetting technique? Access the original article on 17 Ways to Avoid Pipetting Errors, download the Gilson Guide to Pipetting, and read the Nature article showing the effect of sample temperature on pipetting volume. [1, 2, 3]

Resources:
1. 17 Ways to Stop Pipetting Errors From Ruining Your Experiments. Available at: https://bitesizebio.com/344/17-ways-to-stop-pipetting-errors-ruining-your-experiments/
2. Gilson Guide to Pipetting. Available at: https://gb.gilson.com/GBSV/guide-to-pipetting
3. Millet, F., Barthlen, T. Securing accuracy and precision when pipetting hot and cold liquids with Microman®. Nat Methods 4, iii–iv (2007). https://doi.org/10.1038/nmeth1086

#1 — Is it worth doing a PhD? This is a question that probably plagues every research student at some point in their career.

In this episode, we explore 5 important questions you should consider before embarking on a PhD.

Read the full article for more advice before making the difficult decision to pursue a PhD after getting your Master’s degree. [1]

If you are looking for further advice, make sure you check out our article with pointers for PhD students. [2] A good PhD supervisor is worth their weight in gold and finding a good mentor should be a priority. [3] And, if you’re sure that a PhD is the right move for you, then search for PhDs in Biological and Medical Sciences to find the right PhD to suit you. [4] Finally, read our handy article that lists some alternative career options for scientists. [5]

Resources:
1. Is it Worth Doing a PhD After a Master’s? Available at: https://bitesizebio.com/11022/is-it-worth-doing-a-phd-after-a-masters/
2. 10 Do’s and Don’ts for PhD Students. Available at: https://bitesizebio.com/articles/10-dos-and-donts-for-phd-students/
3. Picking an Advisor: The Good, The Bad, and The Ugly. Available at: https://bitesizebio.com/articles/picking-an-advisor-the-good-the-bad-and-the-ugly/
4. Find a PhD. Available at: https://www.findaphd.com/phds/biological-and-medical-sciences/?10gc00
5. Alternative Careers For Scientists. Available at: https://bitesizebio.com/301/alternative-careers-for-scientists/

#100 — Science attracts so many different and quirky personalities that you are bound to have some people you just don’t get along with. Conflicts happen, and there are many strategies you can take to deal with conflict in the lab. But when your lab supervisor is the problem, it can be a big issue for you.

In this episode, delve into the challenges of dealing with difficult lab supervisors. [1] We identify five common problematic personality traits: passive-aggressive, manipulative, unfocused, micro-manager, and negative reinforcement. Listen and explore practical strategies for addressing each type while remaining professional and constructive.

Check out our related article for further advice on dealing with conflicts in a busy research lab. [2]

Resources:
1. Five Types of Difficult Lab Supervisor and How to Handle Them. Available at: https://bitesizebio.com/1540/6-types-of-bad-boss-and-how-to-handle-them/
2. Dealing with Tension and Conflict in the Lab. Available at: https://bitesizebio.com/27373/dealing-tension-conflict-lab/

#99 — So you’ve got your flow cytometry training booked and are one step closer to that precious data.

But if you want to hit the ground running and get some useful data from your samples, there are some little things you'll need to do.

These include reading up on a bit of background theory, understanding the capabilities of different types of cytometers, and thinking about what you want to learn from your experiment.

In this episode of Mentors At Your Benchside, we've compiled cytometry training advice from a core facility manager to help you get the most out of your training sessions and early experiments. [1]

While you are here, why not learn about the components inside cytometers and what they do? [2] Plus, take a step towards fully understanding your data and explore the difference between forward scatter, side scatter, and their corresponding plots. [3]

Resources:
1. Seven Top Tips to Make the Most of Your Flow Cytometry Training. Available at: https://bitesizebio.com/75182/flow-cytometry-training/
2. Demystifying the Flow Cytometry Optics System: A Peek Under the Hood. Available at: https://bitesizebio.com/31638/flow-cytometry-optics-system/
3. Basic Parameters Measured by a Flow Cytometer: What is Scattered Light and Absolute Fluorescence? Available at: https://bitesizebio.com/25310/basic-parameters-measured-by-a-flow-cytometer-what-is-scattered-light-and-absolute-fluorescence/

#98 — Our labs can contain thousands of chemicals, many of which will be past their given expiry date and many of which are expensive to buy and replace.

Replacing them when you don't need to can be a waste of time and grant money. On the other hand, using expired chemicals can lead to failed experiments and confusing results. 

In this episode of Mentors At Your Benchside, learn what types of chemicals are safe to use past their expiry date, which ones you should probably throw out, and why.

Read out the corresponding article for a handy summary table. [1] And while you're diving into reagents, why not check out the different types of antibiotics in molecular biology? [2]

Resources:
1. What Reagents Can You Use Past Their Chemical Expiry Date? Available at: https://bitesizebio.com/74311/chemical-expiry-date/
2. Antibiotics Used in Molecular Biology. Available at: https://bitesizebio.com/10288/antibiotics-used-in-molecular-biology/

#97 — A research interest statement is essential to successfully apply for an academic job. In this episode, we delve into how to craft an outstanding one. [1]

We cover strategies to outline your past, current, and future research in a concise format. We also explain other key elements such as, creating a compelling introduction, detailing research plans, aligning with targeted labs or departments, and writing a strong conclusion. Plus, get tips on personalizing your applications while maintaining clarity and conciseness.

While you're here, check out our related article packed with tips to help you shine at your next job interview. [2] Or, if you're considering working abroad, check out some of its pros and cons. [3]

Resources:
1. How to Write a Killer Research Interest Statement. Available at: https://bitesizebio.com/35364/position-research-interest-statement/
2. How To Shine in a Job Interview. Available at: https://bitesizebio.com/2599/how-to-shine-in-a-job-interview/
3. The Pros and Cons of a PhD or Post-doc in a Foreign Country. Available at: https://bitesizebio.com/7300/the-pros-and-cons-of-a-phd-or-post-doc-in-a-foreign-country-2/

#96 — An appropriate microorganism preservation method can make all the difference in maintaining the viability of your microbial strains because it plays a crucial role in ensuring reproducible results and continuity in research.

In this episode, learn the preservation methods for short- and long-term microbe storage, their pros and cons, and the kit you need to do them.

Check out the corresponding article for a list of helpful references. [1] Learn the main ingredients of cell culture media [2] and the steps you can take to keep mammalian cell cultures healthy. [3]

Resources:
1. How to Preserve Microorganisms: Store Your Cells Better. Available at: https://bitesizebio.com/45159/how-to-preserve-microorganisms/
2. What’s in Your Cell Culture Medium? Available at: https://bitesizebio.com/37034/cell-culture-medium/
3. How To Keep Your Mammalian Cells Happy And Healthy. Available at: https://bitesizebio.com/8146/how-to-keep-your-cells-happy-and-healthy/

#95 — Have you ever accidentally forgotten to add the Kozak consensus sequence to the start of a coding gene? Or forgotten to include the stop codon? Did you clone something, then realize you wanted to tag it with something? Or do you want to add restriction enzymes to your PCR product to make it easier to clone into a plasmid? Overhang PCR may be your answer!

In this episode, we discuss what overhang PCR is, its benefits, and how to perform it in the lab. [1] While you are here, check out our article on TA cloning? [2]

Resources:
1. Overhang PCR: Add Missing DNA Sequences Using Primers. Available at: https://bitesizebio.com/20786/overhang-pcr/
2. Ta-Da! The Magic of Taq and TA Cloning. Available at: https://bitesizebio.com/19709/ta-da-the-magic-of-taq-and-ta-cloning/

#94 — While there are lots of methods to choose from for cleaning up your RNA or DNA samples, for many researchers, phenol-chloroform is the go-to technique.

In this episode, go beyond the basics of how the method works and get expert practical guidance on performing and optimizing it.

Plus, learn the differences between the common solvents, how to check and adjust the pH of the phenol phase, and get tips to reduce the amount of interphase. 

Check out the corresponding online article for a diagram illustrating how you can reduce the interphase. [1] Learn the theory behind the technique in our easy explainer [2] and explore four other ways you can clean up DNA samples. [3]

Resources:
1. Practical Application of Phenol-Chloroform Extraction. Available at: https://bitesizebio.com/3651/practical-application-of-phenol-chloroform-extraction/
2. The Basics: How Phenol Extraction of DNA Works. Available at: https://bitesizebio.com/384/the-basics-how-phenol-extraction-works/
3. Five Ways to Clean Up A DNA Sample. Available at: https://bitesizebio.com/142/5-ways-to-clean-up-a-dna-sample/

#93 — Bioinformatics is an interdisciplinary field that combines mathematics, computer science, physics, and biology to help answer key questions in modern biological sciences research.

In this episode, we’ve got the lowdown on the training you’ll need to pursue this career path, and a handy list of resources to get you started on your learning. [1] Plus, check out our related article on some of the ways that scientists from diverse fields use bioinformatics. [2]

Resources:
1. How to Become a Bioinformatician. Available at: https://bitesizebio.com/38236/how-to-become-a-bioinformatician/
2. Bioinformatics: It’s Not All About Genomics. Available at: https://bitesizebio.com/46495/bioinformatics-its-not-all-about-genomics/

#110 — Comparing two sets of data is a fundamental process in statistical analysis, crucial for drawing meaningful conclusions across various fields. Whether it's for determining the success of an intervention, understanding market trends, or validating scientific research, the need for comparison arises.

This episode delves into the essence of data comparison, focusing on two prevalent statistical tests: the Student’s t-test and the Mann–Whitney U test. [1] Each test comes with its assumptions and applicability, making the choice between them critical depending on the nature of your data.

Read our related article to learn more about comparing multiple datasets. [2]

Resources:
1. Let’s Talk About Stats: Methods for Comparing Two Sets of Data. Available at: https://bitesizebio.com/19298/comparing-two-sets-of-data/
2. Let’s Talk About Stats: Getting the Most out of your Multiple Datasets with Post-hoc Testing. Available at: https://bitesizebio.com/19318/lets-talk-about-stats-getting-the-most-out-of-your-multiple-datasets-with-post-hoc-testing/

#92 — We all need to lyse cells to extract the goodness—our samples—from them.

However, there are many cell lysis methods. Some are harsh, while some are gentle. Some are laborious, while some are easy. Some require dedicated equipment, while some do not. So which one do you choose?

In this episode, we cover eight cell lysis methods for your experiments. [1] For extra information to help you pick a lysis method, check out our article on the different types of cell walls. [2]

Resources:
1. 8 Cell Lysis Methods to Break Cell Walls. Available at: https://bitesizebio.com/13536/cell-lysis-methods/
2. Cell lysis 101: 5 types of cell walls you need to understand. Available at: https://bitesizebio.com/13535/tutorial-cell-lysis-5-types-of-cell-walls/

#91 — Genomes are complex and encode a vast quantity of information. One of their key features is genetic variants—aberrations in the genetic sequence, usually in the form of insertions, deletions, repeats, and translocations of genetic material.

This episode explains the different types of genetic variants, introduces their key features, and gives you some top tips for studying them.

Read the corresponding online article for links to helpful resources and a handy figure. [1] While you're here, check out how to identify protein binding sequences on DNA using chip ChIP-seq [2] and learn about some of the fascinating applications of genome sequencing. [3]

Resources:
1. Genetic Variants Explained. Available at: https://bitesizebio.com/23996/whats-so-important-about-variants/
2. An Introduction To ChIP-seq. Available at: https://bitesizebio.com/13541/an-introduction-to-chip-seq/
3. Decoding the Genome: Applications of DNA Sequencing. Available at: https://bitesizebio.com/27892/decoding-the-genome-applications-of-dna-sequencing/

#90 — Are you confused about the banding pattern of DNA on agarose gels? DNA can take many structural forms depending on its source and how you have isolated and purified it. And those forms, including linear, nicked, closed circled, and supercoiled, all migrate at different rates on agarose gels.

But how do you identify which band corresponds to which structural form? And why do some of these occur during plasmid preps but not others? Listen to this episode to find out. [1]

Since you're here, check out our article explaining how to get more supercoiled DNA from your plasmid preps [2] and learn how alkaline lysis works. [3]

Resources:
1. How to Identify Supercoils, Nicks and Circles in DNA Plasmid Preps using Gel Electrophoresis. Available at: https://bitesizebio.com/13524/how-to-identify-supercoils-nicks-and-circles-in-plasmid-preps/
2. 6 Ways to Show Your Plasmid Preps Some Tlc and Get More Supercoiled Plasmid in Return. Available at: https://bitesizebio.com/13525/6-ways-to-show-your-plasmid-preps-some-tlc-and-get-more-supercoiled-plasmid-in-return/
3. Lab Basics: How The Alkaline Lysis Method Works. Available at: https://bitesizebio.com/180/the-basics-how-alkaline-lysis-works/

#89 — "Achieve more by doing less" sounds like a piece of cheap advice, but there is a lot of wisdom in it.

Research is complicated. You must choose the best questions to ask, techniques, controls, organisms, and equipment, to name just a few things that make up good experiments. With so much to focus on, it becomes harder to do each of these things well.

This episode explains three actionable steps you can take to simplify your research and become more effective at it.

Check out the corresponding online article for links to related resources. [1] and listen to our The Happy Scientist podcast for advice to stay happy, focused, and satisfied in the lab. [2]

Resources:
1. Simplicity in Science: How to Increase your Research Effectiveness by Doing Less. Available at: https://bitesizebio.com/63207/become-an-effective-researcher/
2. The Happy Scientist: Available at: https://thehappyscientist.bitesizebio.com/

#88 — Getting the best out of your in situ hybridizations requires choosing the correct protocol, deciding if sections or whole mount is better, using the right equipment, making fresh buffers, careful planning for all steps, optimizing your probe concentration, and taking the time to get the development step right. In other words, there are a lot of ways in situ hybridizations can go wrong!

This episode walks you through your first in situ hybridization and how to totally nail it! [1] When you've finished listening, why not check out our article on fluorescence in situ hybridization (FISH)? [2]

Resources:
1. How to Totally Nail Your First in situ Hybridization. Available at: https://bitesizebio.com/44651/how-to-totally-nail-your-first-in-situ-hybridization/
2. Do You Know Where Your mRNAs Are? Available at: https://bitesizebio.com/13391/its-10-am-do-you-know-where-your-mrnas-are/

#87 — Selecting the right blood collection tubes for your experiment is crucial. But do you know what tubes to use for which type of blood sample?

In this episode, we cover the nuances of choosing the appropriate blood collection tubes, a choice that hinges largely on whether you're aiming to collect serum or plasma samples. Understand the vital role that different tubes play in either fostering or preventing blood clotting and how these subtle differences can significantly impact the outcomes of your experiments. [1]

Whether you are venturing into the world of hematology microscopy, exploring genetic material, or identifying circulating biomarkers, choosing the correct tube can be a game-changer. Download our handy poster to illuminate the intricacies of plasma and serum tubes and how the type of tube you choose aligns with your scientific objectives. [2]

Resources:
1. Blood Collection Tubes: How to Pick the Correct One. Available at: https://bitesizebio.com/23701/choosing-the-right-blood-collection-tubes/
2. Poster. Available at: https://bitesizebio.com/blood-collection-tube-chart/

#86 — qPCR primer design is a bit of science, a bit of magic, and a little bit of luck. In this episode, we cover the science of qPCR primer design, a cornerstone in conducting successful qPCR or RT-qPCR assays for gene expression analysis. [1] Get top tips and learn why dedicating time to crafting high-quality primers can save your experiment from poor data and resource wastage. [2]

We guide you through utilizing popular tools like NCBI's Primer-BLAST for your qPCR primer designs, alongside introducing alternative online resources to aid in your experimentation journey. [3] Whether you're beginning your research journey or looking to hone your skills further, this episode promises to equip you with the knowledge to navigate the complex yet captivating world of qPCR primer design proficiently.

Resources:
1. A Step-by-Step Guide to Designing qPCR Primers. Available at: https://bitesizebio.com/10041/designing-qpcr-primers/
2. 10 Tips for Consistent qPCR. Available at: https://bitesizebio.com/1118/10-tips-for-consistent-real-time-pcr-rtpcr/
3. NCBI tool Primer-BLAST. Available at: https://www.ncbi.nlm.nih.gov/tools/primer-blast/

#85 — Working with living cells is a tricky business, and tiny fluctuations in environmental conditions can affect their physiology and impact your experiments. Or worse, it can lead to their death. Game over!

Passaging your cells involves removing them from their growth medium to transfer them to fresh vessels with fresh media.

In this episode, learn how to passage adherent and suspension cells while minimizing the impact on them. See the different passaging strategies you can adopt and decide which ones are right for you!

Check out the corresponding online article for handy tables and figures. [1] Learn all about cell confluency and why it matters [2] and explore the critical components of cell culture media. [3]

Resources:
1. How To Passage Cells in Culture. Available at: https://bitesizebio.com/13680/how-to-passage-cells/
2. How Confluent Are Your Cells? A Beginner’s Guide to Measuring Cell Culture Confluency. Available at: https://bitesizebio.com/63887/cell-confluency/
3. What’s in Your Cell Culture Medium? Available at: https://bitesizebio.com/37034/cell-culture-medium/

#84 — Every experiment starts by preparing some buffer solutions. And every buffer solution starts with weighing out some compounds on an analytical balance.

But these essential yet sensitive pieces of lab equipment are prone to measurement drift—meaning you could be weighing out different amounts every time you use one.
 
In this episode, get three easy tips to avoid measurement drift and learn some reliability testing methods to check your lab balance and prepare solutions with confidence.

Check out the full online article here [1] and read our essential guide to cleaning and caring for lab balances. [2] Plus, check out these eight steps you can take to improve lab accuracy and precision. [3]

Resources:
1. 3 Easy Tips for Avoiding Measurement Drift in Analytical Balances. Available at: https://bitesizebio.com/33245/drift-measurements-analytical-balances/
2. How to Clean and Calibrate Your Lab Balance. Available at: https://bitesizebio.com/24193/how-to-clean-and-calibrate-your-lab-balance/
3. How to Measure and Improve Lab Accuracy and Precision. Available at: https://bitesizebio.com/55470/accuracy-and-precision/

#83 — Chemically competent cells are a key resource in molecular biology labs. But do you really understand what is meant by chemically competent?

In this episode of Mentors At Your Benchside, we explain the science behind them and share how you can make your own stocks, saving you money and avoiding nasty surprises when someone forgets to reorder stocks.

Be sure to visit the original article for helpful diagrams and tables, [1] download our free chemically competent cells cheat sheet for a handy print protocol, [2] and learn how to make your own electrocompetent cells. [3]

Resources:
1. How to Make Your Own Chemically Competent Cells. Available at: https://bitesizebio.com/30145/chemically-competent-cells/
2. Bitesize Bio's Chemically Competent Cells Protocol. Available at: https://bitesizebio.com/chemically-competent-cells-protocol/
3. DIY Electrocompetent E. coli. Available at: https://bitesizebio.com/969/diy-electrocompetent-e-coli/

#109 — How do you build a scientific network that gives you the best chance of getting your research funded? How can you identify who to include in your network, and how should you contact them?

This episode explains how to build a scientific network that works for you. We discuss the answers to these questions and provide some examples of collaborations that ended well—and some that didn't.

Check out our online article for additional resources to help get your research funded. [1] If you struggle to convey the impact of your research, you should definitely check out our webinar on this topic. [2] Plus, you can find more direct funding advice from Joel Berry here. [3]

Resources:
1. Personal Advice on Building Your Professional Network. It Takes a Village. Available at: https://bitesizebio.com/77734/professional-network/
2. How to Effectively Communicate Your Research. Available at: https://events.bitesizebio.com/how-to-effectively-communicate-your-1/join
3. How to Become an Expert at Getting Funded. Available at: https://bitesizebio.com/77308/how-to-become-an-expert-at-getting-funded/

#82 — Did you know that most centrifuge accidents result from user error and improper centrifuge care? While proper balancing of samples is important, it is not the only thing you need to be aware of when using your centrifuge. In this episode of Mentors At Your Benchside, we give you 5 tips to help ensure your centrifuge keeps spinning.

Read the full article to get the formula for finding the real maximum speed to use based on a solution’s density. [1] Need to use an ultracentrifuge? These beasts should be respected, but there is no need to be petrified of them; just read our tips on using an ultracentrifuge safely. [2] Finally, for more advice on using centrifuges, be sure to watch the on-demand webinar from Eppendorf on Essentials in Centrifugation to get informed on how centrifuges work, practical tips and tricks, and critical safety information. [3]

Resources:
1. How to Keep Your Centrifuge Alive: 5 Easy Tips. Available at: https://bitesizebio.com/31550/keep-centrifuge-alive/
2. Respect the Ultra. Available at: https://bitesizebio.com/4251/respect-the-ultra/
3. Essentials in Centrifugation - Better Safe than Sorry! (Webinar). Available on-demand: https://events.bitesizebio.com/essentials-in-centrifugation-1/join

#81 — You’re probably aware of the two main types of restriction cloning (sticky-end and blunt-end cloning), but do you know the difference? And do you know how to do both?

In this episode, we share what blunt-end cloning is, how it works, and give you tips and tricks for performing blunt-end cloning.

Visit the original article for detailed diagrams and helpful tables, [1] brush up on how DNA ligase works, [2] and reacquaint yourself with the different DNA polymerases. [3] You can also get help to improve your blunt end ligations, [4] and discover the magic of TA cloning. [5]

Resources:
1. A Beginner’s Guide to How Blunt-End Cloning Works. Available at: https://bitesizebio.com/45131/a-beginners-guide-to-how-blunt-end-cloning-works/
2. DNA Ligation: How it Works & 6 Top Tips.  Available at: https://bitesizebio.com/10279/the-basics-how-does-dna-ligation-work/
3. Get To Know Your DNA Polymerases. Available at: https://bitesizebio.com/24551/get-to-know-your-dna-polymerases/
4. 10 ways to improve blunt-end ligations. Available at: https://bitesizebio.com/19149/7-ways-to-improve-blunt-end-ligations/
5. Ta-Da! The Magic of Taq and TA Cloning. Available at: https://bitesizebio.com/19709/ta-da-the-magic-of-taq-and-ta-cloning/

#80 — Dive into the fascinating world of histology as we explore the basics of Hematoxylin and Eosin (H&E) staining, a cornerstone technique in tissue study. [1]

Whether you're a budding biologist or just curious about cellular structures, this episode of Mentors At Your Benchside is your introductory guide to the history of H&E staining, its mechanism of action, and the structures it effectively stains. Discover how the positive charge of hematoxylin stains nucleic acids blue, and how the negatively charged eosin brings out the pink in proteins.

And, if you want to go beyond hematoxylin and eosin staining, why not download our free histological stains poster and brighten up your lab? It lists common stains, the color they stain tissues, and some useful notes! [2]

Resources:
1. A Beginner’s Guide to Hematoxylin and Eosin Staining. Available at: https://bitesizebio.com/13400/a-beginners-guide-to-haematoxylin-and-eosin-staining/
2. Bitesize Bio's Histological Stains Poster. Available at: https://bitesizebio.com/histological-stains-poster/

#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/

#108 — What should you use to fix your cells? Alcohols or aldehydes? Gluteraldehyde or formaldehyde? And how long will your cells stay fixed?

This episode explains the four main fixatives for histology and cytometry and when to use them. It also provides some practical tips to ensure your fixation works and explains the benefits of combining fixatives.

Check out the corresponding article for links to related resources. [1] To learn more about autofluorescence and the controls you need to check for it, read this article. [2]

Resources
1. 4 Fixatives for Histology and Cytometry. Perfect Your Preservation. Available at: https://bitesizebio.com/22141/fixation-and-flow-cytometry/
2. 5 Controls for Immunofluorescence: A Beginner’s Guide. Available at: https://bitesizebio.com/44370/controls-for-immunofluorescence-a-beginners-guide/

#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/