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Plasmids 101: Visualizing Subcellular Structures & Organelles

Posted by Susanna Bachle on Jun 22, 2017 10:30:00 AM

The same way the human body is made up of organs, cells comprise compartments and structures, called organelles. Take a sneak peak inside a cell with the images from the Allen Cell Explorer (1).

When studying the function of a protein or its role in a disease, researchers often isolate proteins of interest and examine them using biochemical methods thus removing the context of the cell. However, much knowledge about functionality can be gained by understanding the location and transport of the protein within a living cell. Analyzing differences in protein localization and transport between healthy and diseased states can also provide interesting insights into disease mechanisms and protein function.

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Topics: Fluorescent Proteins

How Dry Ice Affects Sample pH & How to Avoid It

Posted by Leila Haery on Jun 20, 2017 10:30:00 AM

We’re always looking for ways to improve our shipment processes. After reading a publication describing how short term storage on dry ice can shift sample pH, we wondered whether or not the dry ice we use to keep viruses frozen during shipment was having an impact on the samples. We therefore devised a few experiments to determine if our tubes were permeable to the CO2 released from dry ice, and whether this affected the pH of our viral samples. Read on to learn how aqueous samples might be affected by dry ice, and specifically how dry ice can affect virus from Addgene.

Bottom line: there’s good news and there’s bad news. The bad news is that some of tubes’ o-rings are, in fact, permeable to CO2 at low temperatures (-80°C) and once in the tube, the CO2 can alter the pH of the liquid sample. The good news is that this effect is reversible and the pH shift can be prevented. Keep this information in mind if you’re planning on shipping something on dry ice or if you’re receiving samples on dry ice - it may prevent you from seeing some unexpected results.

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Topics: Viral Vectors

New Educational Resource: The Addgene Videos Page

Posted by Eddy Page on Jun 16, 2017 10:00:00 AM

Videos can save researchers time by quickly relaying key points about an unfamiliar topic or lab process. Along with our role as a plasmid repository, Addgene aims to be a vital educational resource for scientists around the world. With that in mind, we are very excited to debut our brand new Addgene Videos page!

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Topics: Science Communication

Choosing the B(right)est Fluorescent Protein: Aggregation Tendency

Posted by Guest Blogger on Jun 15, 2017 10:30:00 AM

This post was contributed by guest bloggers Joachim Goedhart and Marieke Mastop from the Section of Molecular Cytology and Van Leeuwenhoek Centre for Advanced microscopy, University of Amsterdam.

The previous two posts in this series described a practical approach to selecting a bright fluorescent protein and a photostable fluorescent protein. In the third post of this series, we will discuss how to select a non-aggregating fluorescent protein.

In the jellyfish Aequorea victoria, AvGFP forms a homodimer. In corals, the red fluorescent proteins form tetramers. In general, fluorescent proteins have a natural affinity and a tendency to form higher order aggregates. This property can be tolerated in some applications (e.g. labeling of cells or tracking promotor activity), but it is problematic in applications in which the fluorescent protein is used as an inert protein module. This is explained in more detail here. There are a variety of methods that can be used to measure your fluorescent protein’s propensity to aggregate. The basics and pitfalls of these experiments are discussed here.

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Topics: Fluorescent Proteins, Choosing the Brightest Fluorescent Protein

Important Considerations When Using AAVs

Posted by Guest Blogger on Jun 13, 2017 10:30:00 AM

This post was contributed by guest blogger Katrina Armstrong, a Neurophysiology Msc Student at the University of Manitoba.

  1. Location, Location, Location!
  2. Failure to Plan (for Storage) Is Planning to Fail
  3. Patience Is Bitter but Its Fruit Is Sweet
  4. The Future?

Need Virus? Check out Addgene's New Viral Service!

I knew little about adeno-associated Viral Vectors (AAVs) before starting my graduate program at the University of Manitoba. Our lab has been utilizing chemogenetics (Designer Receptors Exclusively Activated By Designer Drugs, DREADDs) and optogenetics as tools to investigate the roles of certain cell types in locomotion. We have relied heavily upon AAV vectors to deliver chemogenetic/optogenetic constructs into our cells of interest. Although they have a small packaging capacity, AAV vectors were suitable for our needs for the following reasons:

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Topics: Viral Vectors

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