Latest Posts

All Posts

Plasmids 101: Secondary Nanobody Toolbox

Posted by Beth Kenkel on Feb 27, 2018 9:04:41 AM

Western blots. ELISAs. Immunofluorescence. What do all of these techniques have in common? They all typically require secondary antibodies, frequently of the mouse or rabbit variety. While antibodies certainly aren’t “broken,” their production does require continued animal sacrifice. Could there be an alternative method for immunodetection? Enter the Görlich lab and their anti-mouse and -rabbit IgG secondary nanobodies toolbox. Nanobodies are like tiny antibodies which work just as well, if not better, than antibodies for all of the above listed molecular techniques, but they can also be expressed in bacteria and extracted with common protein purification methods. Read on to learn more about nanobodies and how their structure and function compare to IgG antibodies, as well as how to produce them for use in your lab.

Read More >

Topics: Hot Plasmids, Plasmids 101

Plasmids 101: Walkthrough of Addgene’s Snapgene-Powered Quality Control Process

Posted by Amanda Hazen on Feb 1, 2018 10:07:31 AM

We’ve talked a lot about the quality control process at Addgene by introducing our new sequencing partner seqWell and going into detail about how we use next generation sequencing results to perform quality control on deposited plasmids. We’ve also talked about how our new Snapgene generated maps provide improved feature detection with an easy to use interface. We regularly use Snapgene for our quality control process because of its expansive feature library and useful tools. In this blog, we’ll walk you through how a scientist at Addgene uses Snapgene to confirm the sequence of a plasmid and we’ll highlight some of the new features available on our website through our Snapgene powered maps and sequence analysis tools.

Read More >

Topics: Plasmid How To, Plasmids 101, Plasmid Cloning

Plasmids 101: Inducible Promoters

Posted by Mary Gearing on Jan 18, 2018 9:34:59 AM

Promoters control the binding of RNA polymerase and transcription factors. Since the promoter region drives transcription of a target gene, it therefore determines the timing of gene expression and largely defines the amount of recombinant protein that will be produced. Many common promoters. like CMV, EF1A, and SV40 promoters, are always active and thus referred to as constitutive promoters. Others are only active under specific circumstances. In this post, we’ll discuss inducible promoters, which can be switched from an OFF to an ON state, and how you might use these in your research. Keep a look out for a future post where we’ll discuss repressible promoters.

Read More >

Topics: Plasmids 101

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.

Read More >

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.

Read More >

Topics: Plasmids 101, Fluorescent Proteins

Blog Logo Vertical-01.png

Subscribe to Our Blog