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Introducing an All-in-One CRISPR/Cas9 Vector System for Multiplex Genome Engineering

Posted by Kendall Morgan on Nov 12, 2014 10:54:00 AM

A newly established all-in-one vector construction system for CRISPR/Cas9-mediated multiplex genome engineering is now available thanks to researchers at Japan’s Hiroshima University who described their new tool in Scientific Reports in June.

“The multiplexity is one of the most advantageous properties of CRISPR/Cas9 compared to ZFNs and TALENs,” said Tetsushi Sakuma of Hiroshima University. “However, there had been no systematically established way of making an all-in-one vector for multiplex genome engineering.”

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Topics: CRISPR, Plasmid Kits

Plasmids 101: Common Lab E. coli Strains

Posted by Matthew Ferenc on Nov 7, 2014 9:56:00 AM

This post was updated on Nov 14, 2017.

You've worked hard designing your plasmid – you carefully selected the optimal promoter for your gene of interest, painstakingly cloned into the perfect empty backbone, made sure to add the right tags to your gene, and may have even put a fluorescent protein downstream, separated by an IRES element. You did a lot of work! But let’s take a moment to recognize your little prokaryotic minions that carried out the labor-intensive process of replicating your new plasmid: the Escherichia coli bacteria.

It’s hard to count the number of different commercial strains of E. coli currently available  a quick Google search suggests there are hundreds. This only includes general lab strains designed for subcloning or protein expression. If you were to include customized strains, the number is probably in the thousands! The goal of this article is to provide enough background for you to distinguish the features of any common lab strain and determine whether it is appropriate for propogating your plasmid or carrying out your experiment.

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Topics: Lab Tips, Plasmids 101

Tips for Using FRET in Your Experiments

Posted by Benoit Giquel on Nov 5, 2014 10:41:00 AM

The first time I heard about FRET during a journal club, my guitarist brain automatically thought about the raised element found on the neck of my guitar...not really useful for a biologist you would say. The student was of course talking about the now well-known FRET, aka Fluorescence (Förster) Resonance Energy Transfer, technique which allows the detection of molecules' interactions, modifications or dissociations in situ. Used since the mid-90s, this technique has revolutionised the way we apprehend molecular complexes and is still a very useful tool.   

Like a guitar hero (that I’m not), FRET loves playing “live”. Indeed, FRET was one of the first techniques which enabled the measurement of single molecule interactions in living cells using a microscope. Historically, molecular interactions were detected by indirect means often using probes with the potential to target several molecules. By analogy, it was like pointing out a group of students in a university hall but not knowing if these students know or interact with each other. FRET reduced the scale of our perception about molecular interactions.

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Topics: Imaging, Fluorescent Proteins

Countdown to Halloween @Addgene

Posted by Caroline LaManna on Oct 29, 2014 1:26:27 PM

Halloween is only 2 days away and here at Addgene we couldn’t be more excited! As a small, tight-knit organization we thrive on 3 things: fun, teamwork, and competition. And our Halloween celebration this Friday will combine all of these.

Addgenies have been organizing Halloween festivities for years – searching our photo archives I found pictures back through 2009, though I suspect that our costume contests were indeed occurring earlier but that the photo evidence was destroyed to protect the innocent revelers. Our Halloween costume contests have become an annual tradition in which we choose our teams and start brainstorming months in advance. Our teams comb the news (pop culture events of the past year obviously make for good costumes), science and our plasmid collection (what science has been hot this year and how do you creatively turn it into an outfit?), and of course there’s always nostalgia (old board games and movies can provide great character costumes).

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Topics: Fun, Inside Addgene

How-to: 5 Steps to a Great Panel Discussion

Posted by Joanne Kamens on Oct 22, 2014 9:57:29 AM

This post was originally published on LinkedIn. Follow Addgene on LinkedIn for repository news and updates.

Panel discussions are becoming a ubiquitous format for events and sessions in scientific conferences. They can result in lively discussions with both panelists and audience fully engaged. They can also be dull and painful to sit through (imagine the entire audience playing Candy Crush or reading email). My advice is always "don't do it unless you can do it well" – even the smallest program. I don't mean have fancy food or a fantastic view (these are nice if you can afford them). I am talking about making sure the audience is interested, educated and talking as a result of the content. Here are some of my tips to organizing and running a memorable panel discussion.

Check out Joanne's Reddit AMA

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Topics: Career

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