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Plasmids 101: NGS Quality Control for Pooled Libraries

Posted by A Max Juchheim on Oct 26, 2017 9:59:02 AM

In addition to single plasmids, Addgene also distributes pooled plasmid libraries containing hundreds, thousands, or even a million plasmids. These libraries are some of Addgene’s most exciting and versatile offerings! We recently re-amplified our distribution stock of the Brunello Human gRNA library, and we thought it would be a good time to talk about the amplification and verification processes we use to ensure high-quality library distribution. You can also use these tips as a starting point when you need to amplify a library for your own experiments.

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Topics: Inside Addgene, Plasmids 101

Plasmids 101: Monitoring Cell Mobility Using Fluorescent Proteins

Posted by Benoit Giquel on Aug 15, 2017 9:24:39 AM

In complex metazoans, rapid cell division and large scale cell mobility are essential processes during embryonic development. These are required for a growing organism to make the complicated transition from a clump of cells to a fully differentiated body. In contrast, these dynamic processes are largely absent in adult organisms, where tissues structures are more stable and local movements predominate (e.g. a basal progenitor cell migrating to the epithelium). At this stage, only cells from the immune system show wide scale mobility with movement from the bone marrow and other lymphoid organs to specific tissues where they can scan for any signs of danger. In this post we’ll focus on how fluorescent proteins can and have been used to monitor cellular movements in the immune system. The techniques used here could be adapted to studying other systems in which there is large scale cellular movement throughout an organism.

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Topics: Plasmids 101, Fluorescent Proteins

Plasmids 101: An Inside Look at NGS Plasmid Quality Control

Posted by Amanda Hazen on Aug 3, 2017 8:49:45 AM

All plasmids coming through Addgene’s doors are now verified by next-generation sequencing (NGS) to provide you with more data. For most plasmids, we are now able to confirm the full insert and backbone sequence. In order to manage and interpret these data, we’ve updated our quality control workflow.  Here’s a peek at how our process has improved now that we’re backed by the power of NGS!

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Topics: Inside Addgene, Plasmids 101

Plasmids 101: Introduction to FRET

Posted by Jason Niehaus on Jun 27, 2017 9:03:20 AM

Imagine being able to determine whether two proteins are within 10 nanometers of each other, or measure the tension in the helical structure of spider silk, or the activity of a protein in a synapse. What kinds of tools enable us to measure these properties, and what fascinating experiments could push these tools even further? All of these things can be done using FRET! Read on to find out more about this amazing imaging technique and find further tips for using FRET in your experiments here.

 

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Topics: Plasmids 101, Fluorescent Proteins

Plasmids 101: Fluorescent Protein Timers

Posted by Tyler Ford on May 4, 2017 10:30:00 AM

Even before fluorescent proteins (FPs) came into wide use, there were a variety of ways to monitor cell, organelle, and protein localization. For instance, you might dye your cells and look at them under a microscope, fractionate samples to isolate particular organelles and their contents, or perform in situ hybridization experiments. In many cases fluorescent proteins have usurped old methods or complemented them in ways that make them much easier. A special class of FPs, the FP timers, add an entire new dimension to monitoring localization; using FP timers, researchers can look at a single image of a cell and understand how protein localization changes over time.
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Topics: Plasmids 101, Fluorescent Proteins

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