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ICYMI: Addgene’s Plasmids 101 eBook

Posted by Marcy Patrick on Oct 2, 2014 1:25:00 PM

We'd like to thank all of the contributing Addgenies that made this eBook possible: Melina Fan, Matthew Ferenc, Larissa Haliw, A. Max Juchheim, Caroline LaManna, Margo Monroe, Kendall Morgan, Jason Niehaus, Marcy Patrick, Lianna Swanson, Julian Taylor-Parker

We'd also like to thank our guest contributor: Gal Haimovich of for helping us explain why things glow!

Addgene's Plasmids 101 eBook is here: Enjoy more time developing clever experiments and less time researching basic plasmid features – download the Addgene Plasmids 101 eBook!

Our goal was to create a one-stop reference guide for plasmids. We’ve combined our Plasmids 101 blog posts from the last year with some additional resources to create one downloadable PDF you can save to your desktop for easy reference. Highlights include our guide to fluorescent proteins, information about promoters and ORIs, and tips for naming your plasmids.  

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Topics: Plasmid Elements, Lab Tips, Plasmids 101

Plasmids 101: Multicistronic Vectors

Posted by Melina Fan on Sep 9, 2014 4:20:00 PM

Co-expression of multiple genes is valuable in many experimental settings. To achieve this, scientists use a multitude of techniques including co-transfection of two or more plasmids, the use of multiple or bidirectional promoters, or the creation of bicistronic or multicistronic vectors. Unlike promoters which will create unique mRNA transcripts for each gene that is expressed, multicistronic vectors simultaneously express two or more separate proteins from the same mRNA. We've discussed promoters before so in this blog post we’ll cover basics of multicistronic vectors: why they are useful, how they work, and how to get started with them.

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Topics: Plasmid Technology, Plasmids 101

Plasmids 101: How to Verify Your Plasmid

Posted by Lianna Swanson on Aug 28, 2014 11:34:00 AM

Congratulations, you have a plasmid expressing your gene of interest (YGOI) and are ready to dive into your functional experiments! Whether you’ve cloned the plasmid yourself or obtained it from a colleague down the hall, it is always a good idea to take some time to confirm that you are working with the correct construct, and verify that the plasmid you received matches the expected sequence. Here at Addgene, we use NGS-based quality control to confirm the sequence of all the plasmids we distribute. This method is time-intensive, so we recommend two other methods for quick plasmid verification: Sanger sequencing and diagnostic restriction digest.

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Topics: Plasmid How To, Lab Tips, Plasmids 101

Plasmids 101: Viral Vector Elements

Posted by Marcy Patrick on Jul 17, 2014 3:09:00 PM

The use of viral vectors in research is beneficial for a number of reasons, including but not limited to: helping to get difficult-to-deliver DNA into mammalian cells, increasing the efficiency of gene transduction, allowing for control over which cells are infected through viral pseudotyping, and ease of vector cloning and modification. At the most basic level, viral vectors consist of a viral genome that has been adapted into a plasmid-based technology and modified for safety through the removal of many essential genes and the separation of the viral components. Read on for a brief description of the viruses used to make these vectors as well as a table defining the major elements found within the plasmids comprising the viral vector systems.

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Topics: Plasmid Elements, Plasmids 101, Viral Vectors

Plasmids 101: Luciferase

Posted by Jason Niehaus on Jun 24, 2014 11:59:00 AM

Luciferases are a class of enzymes capable of catalyzing chemical reactions in living organisms resulting in the emission of photons. The most familiar bioluminescent organism for most people is the firefly (Photinus pyralis) and perhaps not surprisingly it is also the most commonly used bioluminescent reporter. This beetle emits a yellow-green light with a peak emission at 560nm. Shortly after the initial article describing the cloning of firefly luciferase was published in 1985, several studies utilized luciferase as a genetic reporter in plant and mammalian cells. Luciferase assays have since become a gold standard in gene expression analysis and a luciferase gene (one of many available to choose from) is now a common feature in reporter plasmids. 

Learn How Luciferase Can Be Used In Concert with Fluorecent Proteins in Nano Lanterns

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Topics: Plasmid Technology, Plasmids 101

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