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Opto-Nanobodies: Using Light to Manipulate Cell Signaling and Protein Purification

Posted by Beth Kenkel on Nov 19, 2019, 9:08:50 AM

It’s time to choose your own protein purification adventure. You want to purify your favorite protein (YFP). You have two options: 

Option #1: Affinity tag purification

You tag YFP and use an affinity column for purification. After binding YFP to the column, you wash several times to remove non-specific proteins, and then elute YFP. 

Option #2: Opto-Nanobodies (OptoNBs) purification

You skip adding a tag to YFP and instead use OptoNBs. You fill a column with OptoNB coated beads and wrap the column with blue LED lights. When you switch off the lights, OptoNBs bind YFP and non-specific proteins flow through. To elute YFP, you turn on the blue lights.

Which option do you choose? 

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Topics: Optogenetics, Other Plasmid Tools, Plasmids

Behind-the-scenes of the Isolation of the Thermostable IgnaviCas9 From a Yellowstone Hot Spring

Posted by Christina Mork on Nov 12, 2019, 9:00:00 AM

In 2008 the Quake Lab at Stanford University became interested in exploring biological dark matter – large tracts of the microbial tree of life that remained unexplored. Using new single-cell sequencing approaches, the lab was able to eliminate the need for axenic (pure) laboratory cultures to study these microbes. From 16S rRNA sequencing, hot springs were known to be diversity hotspots containing abundant biological dark matter and so the lab organized a sampling trip to Yellowstone National Park (YNP).

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Topics: CRISPR, Cas Proteins

What Good Citizenship Can Do for Reproducibility in Science

Posted by Guest Blogger on Nov 7, 2019, 9:18:13 AM

This post was contributed by Deborah Sweet, Vice President of Editorial at Cell Press.

Almost everyone who works in a lab struggles with reproducibility at some point.

Usually it comes up when a researcher decides on a new project and begins by trying to reproduce someone else’s result. Then, they hit trouble. The experiment won’t work. Even if it does, they don’t get the same result. So, then they end up investing time that they thought would be moving forward instead trying just to get going. It’s like being stuck in jail in Monopoly—you keep rolling the dice and not moving while all your friends are racing around the board. Eventually, you get lucky and manage to escape, but you’ve lost a lot of time.

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Topics: Scientific Sharing, Reproducibility, Scientific Publishing

Choosing a CRISPR Nuclease: Site Accessibility, Specificity, and Sensitivity

Posted by Andrew Hempstead on Nov 5, 2019, 8:28:59 AM

In January 2016 we first published a blog post titled: Which Cas9 Do I Choose for My CRISPR Experiment? The three years flew by, but since then, scientists have adapted CRISPR nucleases for many more specific research needs. In this update, we will focus on the most recent advances and how some of these variants may be appropriate for your specific research question.

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Topics: CRISPR, Cas Proteins

Six Spooky Science Stories and Halloween at Addgene

Posted by Jennifer Tsang on Oct 31, 2019, 9:06:51 AM

Plasmid pumpkins, team costumes, and spooky science stories. Yes, please!

At Addgene, we love finding ways to incorporate science themes into our fun activities and Halloween is no exception. For the past 10+ years, Addgenies have formed teams, chosen a theme for the costume, and did their best to claim the costume contest trophy. We’ve had teams dressed up as plasmids, Blugene, and the Addgene ping pong table.

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Topics: Inside Addgene, Addgene News

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