“Viable But Non-Culturable (VBNC)”: Zombies of the Bacterial World

“Viable But Non-Culturable (VBNC)”: Zombies of the Bacterial World

Imagine that you want to test the efficiency of an antimicrobial treatment in inhibiting a certain bacterial pathogen. As part of the experiment, you expose the bacteria to the treatment and monitor the cultivability of the microorganism by counting the number of colony forming units (CFU) formed on culture media. If the microorganism is sensitive…

8 Tricks to Improve Your Negative Staining of Membrane Proteins

8 Tricks to Improve Your Negative Staining of Membrane Proteins

Negative staining of proteins is a versatile tool for structural biology. The sample preparation protocol is simple: the sample is embedded in a heavy metal stain that gives rise to increased specimen contrast. Thus, negative staining is a very convenient method to assess sample homogeneity, formation of macromolecular complexes, or quality of protein preparation. Conventional…

4 Important Considerations for Your Cell Lysis

4 Important Considerations for Your Cell Lysis

You’ve cultured your cells and completed your treatments, now it’s time to harvest them and proceed to the downstream effects. Cell lysis is the crucial stage that determines if your experiment has a chance of producing the data that you have been waiting for. Part of the starting biological material is inevitably lost on each…

Ten Tips for Pipetting the 384-Well Plate

Ten Tips for Pipetting the 384-Well Plate

I was so excited to start using 384-well plates for my assays. With so many wells, these plates are useful for testing many conditions in parallel, as required in ELISAs, siRNA library screens, and drug treatment dilutions. However, I quickly learned that pipetting in these plates is more complicated than I thought. This article contains…

Need a Few More Hours in the Day?  Here Are Some Timesaving Tips

Need a Few More Hours in the Day? Here Are Some Timesaving Tips

Are you constantly looking for ways to squeeze more lab hours out of the day? Here are some timesaving tips that can help. Figure out Where Your Time Goes Not sure where you are losing those valuable hours? Use a website like Toggl to help you keep track of your time. This will make it…

Don’t Let Bubbles Burst Your Experimental Excitement

Don’t Let Bubbles Burst Your Experimental Excitement

Bubbles isn’t just the name of my favorite cartoon character from Power Puff girls, or just the best activity for a kid to play with, in general. In my adult world, they stand for a whole lot more, but can still cause extreme emotions. At the lab bench, seeing bubbles brings happiness or sadness depending…

Water your choices? Understanding Types of Water in the Lab

Water your choices? Understanding Types of Water in the Lab

If you are working in a scientific laboratory, it is very important to be aware of the various types of water available, because the purity may not be acceptable for your specific experimental application. In most labs, there are generally two types of water piped in to the sinks: Industrial Water Industrial water is non-potable…

Outsourcing Research:  Should Your Experiment Spend Some Time Away from You?

Outsourcing Research: Should Your Experiment Spend Some Time Away from You?

As a researcher, it’s satisfying to manage your own projects and do the bench work yourself. After all, if you don’t have experience with a technique, you’re usually expected to figure it out (with or without direct supervision). In some situations, dealing with difficult molecular techniques is simply part of the job description. The scientific…

Three Steps for Setting up a Drug Screening Assay

Three Steps for Setting up a Drug Screening Assay

An anti-cancer drug or antibody drug conjugate (ADC) screening assay is the first step to establish the utility of a drug candidate in killing cancer cells. Nevertheless, these assays are time consuming and tedious. The purpose of this article is to make things easier when you are required to set up these in vitro screening…

Optimize Bacterial Protein Expression by Considering these 4 Variables

Optimize Bacterial Protein Expression by Considering these 4 Variables

So, you have successfully cloned your gene of interest and are eager to purify buckets of protein. No matter your eventual application—kinetic experiments using a SPR instrument, structural analysis using X-ray crystallography, or any other experiment—you’ll need to express your protein first.  Now, it’s time to put your expression plasmid into E. coli and get…

Making the Most of Quiet Days in the Lab:  From Gloomy to Glorious

Making the Most of Quiet Days in the Lab: From Gloomy to Glorious

It’s Monday morning. You arrive in the lab armed with a large coffee and feeling rested after a non-lab weekend. You check your email and calendar and peek into your PI’s office. Today will be a rare non-experimental day, a day that some love and others dread: a day to clean up and get ready…

Four Tips for Working with Human Clinical Samples

Four Tips for Working with Human Clinical Samples

While using human clinical samples in your research can provide robust and heterogeneous results applicable to larger portions of the population, working with these samples presents its own set of challenges. Here are some tricks I have learned to help isolate and grow your cells of interest while eliminating stromal, blood, or other undesired contaminants….

SPUD’s Your Bud When it Comes to Sensitive qPCR

SPUD’s Your Bud When it Comes to Sensitive qPCR

There’s piloting a brand new technique for the first time. Then, there’s jumping through hoops trying to get an established lab technique to work. The former, in contrast to the latter, is expected to be fraught with hardships. Yet troubleshooting an old lab technique that isn’t working anymore, is frustrating at a whole new level….

Defeat RNAse Contamination Using Bleach in Your RNA Agarose Gel

Defeat RNAse Contamination Using Bleach in Your RNA Agarose Gel

So, you’ve extracted your precious RNA and want to check its quality on a gel. Conventionally, you would run a formaldehyde gel, which is messy and requires a lot of prep. Plus, it is a huge undertaking in terms of time (and money) if all you want to do is just check the quality of…

How to Start Using Coding to Automate Image Analysis Part 2: Batch Processing Multiple Images

How to Start Using Coding to Automate Image Analysis Part 2: Batch Processing Multiple Images

In Part 1 of this article, I introduced you to using code for basic image manipulation in ImageJ and working with the command recorder to expand your coding vocabulary. I covered how to make a simple macro, how to edit it and then save it to be run again another time. If you skipped the…

How to Shut Off Background Lac Expression in LB

How to Shut Off Background Lac Expression in LB

Here’s a tip that you may find useful if you are expressing proteins in E.coli using a lac promoter-based expression system, e.g. pET, in LB medium (L-broth). Lac expression systems are typically induced in the lab using IPTG (isopropyl-beta-D-thiogalacto- pyranoside), which is a non- hydrolyzable analog of lactose, the natural inducer of the lac operon….

5 Tips on Vector Preparation for Gene Cloning

5 Tips on Vector Preparation for Gene Cloning

One of the most crucial steps in any cloning procedure is the preparation of the vector. Get it wrong and your chances of success will be drastically reduced. The overall aim for a good vector preparation is to obtain a fairly concentrated stock of undamaged, fully digested plasmid DNA that is free from contaminants. Missing…

Microbiology 101 by Aunt Yersinia

Microbiology 101 by Aunt Yersinia

We are pleased to announce that the famous maid of microbiology, dear old Aunt Yersinia, has agreed to start writing a microbiology and molecular biology advice column for Bitesize Bio. She will be free to answer your most pressing questions sent to:  auntyersinia@bitesizebio.com By way of introduction, Aunt Yersinia is bestowing 8 spores of knowledge garnered…

Tips for Heating up Agar in the Microwave

Tips for Heating up Agar in the Microwave

One of our readers posted the following question to us and we decided to pass it along to everybody’s favorite microbiology expert, Aunt Yersinia: For one year I am working in different research laboratories, after I got from school. I keep wondering why EVERYBODY is using pre-made Agar solutions for pouring plates, and EVERYBODY is…

Molecular Cooking: How to Apply Science in the Kitchen

Molecular Cooking: How to Apply Science in the Kitchen

Rumour has it that many people who work in a lab enjoy cooking. And, when asked by a non-biologist family member how they spend their days in the laboratory, molecular biologists might answer that doing experiments is similar to cooking. Apart from the organisational parallels that can be drawn between following a protocol and a…

Quick and Dirty Screening for Cloned Inserts

Quick and Dirty Screening for Cloned Inserts

For identifying positive clones from a plasmid cloning procedure, the routine of performing a mini-prep and then checking the putative clones by restriction digestion is most commonly used. Of course, if you need to screen a large number of clones, another option is a colony PCR to identify positives, followed by restriction digests to confirm….