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Multiplex CRISPR Gene Editing: What’s It For?

The RNA-guided nature of CRISPR nucleases (e.g., Cas9, Cpf1) makes them highly amenable to multiplex applications, including multi-gene knockout and large-scale genomic modifications. This article — part 1 of a two-part series on multiplex CRISPR gene editing — discusses these applications and outlines practical considerations for multiplex CRISPR gene editing experiments. New to CRISPR gene…

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Nodulation: What Is It and Why Is It Important?

Plants are incredibly organisms. Not only do they provide atmospheric oxygen, but, in the case of legumes, they can transform atmospheric nitrogen gas to ammonia, which can then be consumed by humans. How does this happen, you ask? It’s all thanks to bacteria and the process of nodulation. Deep Breaths Nitrogen is an incredibly important…

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What the HEK? A Beginner’s Guide to HEK293 Cells 

If you’re at all familiar with cell culture, you’ve probably heard of HEK293 cells. HEK293 are one of the most commonly used cell lines in laboratories across the globe. But what are they? Why should you use them? And what does the ‘293’ mean? Are HEK293T the same thing? Read on for answers to these…

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Successful Start: Tips for Establishing Organoids

Organoids are a developing star of research. They can be grown to represent the majority of mammalian organs and have a wide range of possible applications. More realistic than simple monolayer cell culture, they offer an in-between step that reduces the need for animal models and simplifies (although doesn’t completely remove) ethics paperwork. Once you’ve…

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DIY Centrifugation-Based Purification of cfDNA

There are many reasons you may want to study circulating, cell-free DNA (cfDNA) such as non-invasive prenatal testing to generate a molecular karyotype of an unborn fetus or for use in cancer to detect, diagnose and monitor the disease. Qiagen’s QIAamp circulating nucleic acids extraction kit is consistently cited in the scientific literature as the…

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Making a Mark: How to Set up Your Wound Healing Assay 

Are you wanting to study cell migration and need a way to quantify it? A wound healing assay is one of the earliest and simplest in-vitro assays developed to study cell migration. Collective cell migration is a characteristic behaviour seen in wound repair, cancer invasion, cancer metastasis, immune response, angiogenesis and embryonic morphogenesis. Depending on…

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What’s The Problem With Ampicillin Selection?

Resistance to the antibiotic ampicillin is commonly used as a selection marker for plasmids in gene cloning and protein expression in E.coli and other bacteria. While it can be incredibly useful tool, there can be problems using this selection marker that you need to be aware of if if you plan on using it. This…

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Should You Switch from Wet to Dried Blood Samples?

A Spot of History Most of the biomedical methods used started as a curiosity. Then the one-off gains a limited use, the technology then progresses until its use becomes widespread. Just think about the arch from the curious polished glass spheres, used by Antony Levnhook to look at animalcules, to modern microscopes. The same story…

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How to Preserve Microorganisms: the long (and short) game

Given the importance of microorganisms as cell factories in biotechnological applications and as model organisms for the study of various biological processes, the preservation of microorganisms plays a key role in ensuring reproducible results and continuity in research. Maintaining a library of microbial stocks also enables microorganisms to be easily stored and retrieved, as compared…

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A Beginner’s Guide to How Blunt-End Cloning Works

Blunt and sticky might sound dull and dirty but knowing how these different cloning methods work is important when choosing which method to use. Here we give you a guide to how blunt-end cloning works. Blunt-end cloning is the cloning of DNA fragments containing no unpaired bases at the 5’ and 3’ prime ends (i.e.…

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Antibiotics Used in Molecular Biology

Antibiotics are used in a wide range of techniques in molecular biology including molecular cloning and are important for treating pesky mycoplasma contamination in cell cultures. They can also be used to maximize your plasmid yeilds by reducing protein synthesis, in certain circumstances. The aim with this post is to provide an easy reference to…

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Top Tips for Troubleshooting In Vitro Transcription 

The Phobia of RNases My first experience of troubleshooting in vitro transcription came when I was synthesizing RNA In-Situ Probes for the first time. A lab mate ominously warned me that I had just returned to lab after a bout of flu and that meant I’m a walking talking factory of RNases. I went ahead…

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Dead Useful: CRISPR-Cas9 Epigenome Editing

Given the rapid pace with which genome editing technologies have been developed and adopted, it comes as no surprise that the original CRISPR-Cas9 system has been successfully modified by some very clever scientists. No longer are we limited to the ‘simple’ case of editing the genetic sequence of your biological system of choice, but there…

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DNA Extraction for Next Gen Sequencing

The advent of Next Gen Sequencing (NGS) has been truly amazing. One of the marvels that is often overlooked is how advances in DNA extraction technology have helped streamline NGS workflows. The original standard – phenol/chloroform extraction – is not well suited to the automated nature of today’s sequencing workflows (though with the emergence of…

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Faster Ligations: PEGing down the Secret

Overnight ligations are inconvenient — especially when they fail. Luckily, there’s a straightforward way to faster DNA ligations. This article highlights the secret ingredient to faster ligation reactions and offers some tips and caveats on its use. For a general overview of DNA ligations, see here and here. Buy a Quick Ligation Kit The most…

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Spinning Around: Tips and Tricks for Using Centrifugal Filters

One of the most widespread protein laboratory accessories are the MWCO (molecular weight cut-off) centrifugal filters which are commonly used for concentrating protein, as well as DNA. They are available commercially with different cut-offs including 3kDa, 30kDa, 50kDa, 100kDa, and so on. These little devices are expensive and hence demand proper usage and care to…

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Staying Alive: Tips for Air-Liquid Interface Cultures

What Is Air-Liquid Interface Culture? Long gone are the days where scientists had to rely on 2D cultures of immortalized cell lines to learn principles of human biology. Today, we have a variety of cell culture systems that come closer than ever before to mimicking the structure and function of our body’s organs. One example…

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