Latest Posts

All Posts

Plasmids 101: Protein Expression

Posted by Alyssa Cecchetelli on Jun 7, 2018 9:17:55 AM

The central dogma in molecular biology is DNA→RNA→Protein. To synthesize a particular protein DNA must first be transcribed into messenger RNA (mRNA). mRNA can then be translated at the ribosome into polypeptide chains that make up the primary structure of proteins. Most proteins are then modified via an array of post-translational modifications including protein folding, formation of disulfide bridges, glycosylation and acetylation to create functional, stable proteins. Protein expression refers to the second step of this process: the synthesis of proteins from mRNA and the addition of post-translational modifications

Read More >

Topics: Plasmid How To, Plasmids 101

Plasmids 101: Repressible Promoters

Posted by Mary Gearing on May 10, 2018 9:15:54 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 T7, CMV, EF1A, and SV40, are always active and thus referred to as constitutive promoters. Others are only active under specific circumstances. In a previous post, we discussed inducible promoters, which can be switched from an OFF to an ON state, and how you might use these in your research. Today, we’ll look at repressible promoters, which can be switched from an ON to an OFF state, as well as repressible binary systems commonly used in Drosophila.

Read More >

Topics: Plasmids 101

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. When you're done with this post, check out our follow up post on repressible promoters.

Read More >

Topics: Plasmids 101

Blog Logo Vertical-01.png
Click here to subscribe to the Addgene Blog
 
Subscribe