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Posts Tagged ‘Tech Tips’

The Essential Guide to In-Cell Westerns

In-cell Westerns are a powerful technique that has enhanced how researchers analyze protein expression levels and signaling pathways within fixed cells. Learn about their primary advantages, applications, and some of the best tech and products to perform them.

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What is PCR? The Beginner’s Guide

If you need to copy, sequence, or quantify DNA, you need to know about PCR. Read our guide to the PCR process, and discover tips to help you avoid the most common PCR pitfalls.

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Cloning Methods: 5 Different Ways to Assemble

Over the past few decades molecular biologists have developed procedures to simplify and standardize cloning processes, allowing vast arrays of artificial DNA structures to be more easily assembled. Are you familiar with all the cloning options out there? Let’s look at five different cloning methods you can use to get your construct. At the end…

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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…

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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…

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Why Isn’t My Culture Growing? The S-Curve Explained

Whether you work with human cell lines or microbes, their growth is governed by the same principles. I invite you to learn about something that lies at the base of any work with cell culture, whether cells have circular or linear chromosomes: the S-curve of the population growth. The length of each phase depends on…

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Why You Should Use Cas9 Ribonucleoprotein Transformation for CRISPR Genome Editing

Imagine directly creating a mutation at (almost) any site in your target genome instead of screening thousands or millions of random mutants! The CRISPR/Cas9 system does just that. In its traditional form, this forward genetics approach takes 7 steps from start to mutated genome. However, there is a way to obtain your designer genome in…

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Alphabet Soup for Bacteria!

In its simplest form, a bacterial growth medium is designed to support the growth of bacteria. Depending on which bacteria you want to culture, you may have a range of different media to choose from, each containing a rather unique blend of sometimes surprising (and odd!) components! In this article, I will take you through…

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The Good, the Bad and the Expensive of Whole Genome Sequencing

Whole Genome Sequencing (WGS) is still very cutting edge, sequencing technology and while there are a lot of perks to using it, there are also a few drawbacks. The good, the bad and the pricey are outlined below to help you navigate when it’s worth using WGS! Whole Genome Sequencing: The Good Lots of Data…

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What to Do During That Awkward One-Minute Spin

We’ve all been there. Twiddling our thumbs. Staring off into space. Pacing back and forth. This is the dreaded one-minute spin. If you’ve dabbled in molecular biology, you’ve likely encountered this awkward time. Not exactly enough time to actually do anything else, but when you’ve got nothing to do but wait, one minute seems like…

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Post-sorting Checks and Measures

In my previous article I discussed steps you can implement to ensure that a sample is ready for cell sorting. But now it’s time to make sure the sort worked. Here are a few sorting checks and measures to ensure that all’s well that ends well. Post-sorting Checks and Measures Re-evaluate Your Catch Tubes Sorting…

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Introduction to DREADDs – Control Over G Protein Coupled Receptor GPCR signaling

Gee, Protein, What Do You Do? Manipulation of a system under investigation is the backbone of experimentation. A new tool called Designer Receptors Exclusively Activated by Designer Drugs (DREADDs) allows us to hijack cell signaling and study cell function within living organisms. Like its cousin technique, optogenetics, DREADD technology uses a viral vector to introduce…

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PCR Pitfalls: The Devil is in the Details

PCR was actually one of the first lab techniques I learned as an undergrad. Despite being sometimes labeled as a pretty basic lab skill, PCR doesn’t always work as expected. This “fickle” success is due to small details or hidden hazards within the PCR workflow that can cause your seemingly uncomplicated experiment to fail.  This…

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The 3 Most Common Flow Cytometry Fallacies

Flow cytometry is fast evolving from a method only revered by immunologists, to one used by nearly every biological specialty. It’s pretty much my favorite tool. Unfortunately, as with most lab techniques, much of flow cytometry is taught on the job without a lot of standards. And too often bad habits are passed along like…

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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…

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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…

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Mastering the Art of Growing THP-1 cells

Tissue culture can sometimes seem like a black art. Too careful—your cells go down. Not careful enough—your cells go down. A butterfly flutters its wings in the middle of the Atlantic Ocean—your cells go down. It’s annoying, it’s frustrating, and there are times (and I’m speaking from personal experience here) that you’ll end up chucking…

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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…

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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…

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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.…

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How to Clean and Unclog Your HPLC Column

In my last article, I discussed how to best keep your lab’s HPLC running smoothly. However, even the best-maintained HPLCs and columns need periodic cleaning. Today, I’ll describe how to identify and troubleshoot a clogged HPLC column. Columns Are Finite First of all, it’s important to realize that columns do have a finite lifetime. The…

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DNA Shuffling Like a Pro

DNA shuffling uses PCR technology in a very creative way. It allows you modify your protein to make a new protein you want. You can evolve proteins in microcentrifuge tubes on your very own lab bench. Isn’t that fantastic? DNA shuffling is also a very powerful technique for directed molecular evolution. W. Stemmer first used…

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Under Pressure: Tips for Keeping Your HPLC Up and Running Properly

If you’re anything like me, your biggest lab fear is working with expensive equipment prone to damage. HPLC is a wonderful tool, capable of separating, identifying, and quantifying a vast array of compounds, but it requires an attentive scientist to properly handle and maintain each component. In this article I’ll describe a few basic handling…

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How to Cherry Pick Your Primary Antibody

How do you pick which antibody you should use in your assay? If you’re starting a new assay and need an antibody for the job, then selecting a new antibody from the plethora available could be high up on your to-do list.

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How to Plan a Restriction Cloning Experiment In Silico

Restriction cloning, at its core, is quite simple. You simply cut the target vector and insert with the same enzymes, clean digested vector and insert up, ligate the two together, transform the ligated vector and insert into bacteria, and then screen. While getting each of the steps correct can be a bit of a hassle,…

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Working in a Cold Room Without a Parka?

Have you ever needed to work in a cold room for a long period of time? For example, if you need to dialyze or purify a protein of interest that is temperature sensitive, working in a 4°C cold room might be the only way to accomplish the work. Well, you are in luck. I dislike…

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Six Steps to Successful Mouse Genotyping

An essential step in mouse breeding is genotyping them to determine the genotype of every mouse in the litter. It is also useful to differentiate between various groups of experimental mice if any confusion arises. When genotyping, you will be hunting for the specific gene that you want your mice to have or a genetic…

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Quick and Easy Automatic Cell Counting

Are you wondering how on earth you’re going to count thousands of cells across a stack of images? Well, I’m going to show you a simple method for automatic cell counting with ImageJ. For those of you unfamiliar with ImageJ, it’s a popular image processing program that runs on Mac, Windows, and Linux. Assuming you…

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A Beginner’s Guide to Tag-Removing Proteases

Every biochemist is familiar with proteases. More often than not, proteases cause a lot of anxiety. To this end, a lot of research has been done in developing techniques to prevent the activity of proteases. But some of these proteases can be the good guys too! For example, you can use them to separate your…

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