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Choosing Your Fluorescent Proteins for Multi-Color Imaging

Posted by Guest Blogger on Oct 9, 2014 11:00:00 AM

This post was contributed by Kurt Thorn of the Nikon Imaging Center at UCSF.

A common requirement for live cell imaging experiments is the ability to follow multiple fluorescently tagged species simultaneously. To do so with fluorescent protein labels requires multiple fluorescent proteins whose excitation and emission spectra differ sufficiently for them to be imaged in distinct fluorescent channels on the microscope. With the proliferation of fluorescent proteins in recent years, there are many fluorescent protein combinations that can be imaged together, but this also means that the choice of fluorescent proteins requires some thought.

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Topics: Lab Tips, Fluorescent Proteins

Future of Research Conference - Remarkable Opening Session

Posted by Joanne Kamens on Oct 7, 2014 11:00:00 AM

 

It was with a sense of relief that I attended the amazing, postdoc-driven Future of Research Conference at Boston University last week.  This fantastic city-wide effort was led by two amazing postdocs, Kristin Krukenberg and Jessica Polka from Harvard Medical School and a long list of supporters from all of the research centers in Boston.  I was relieved because it finally feels to me like the postdocs are taking some steps to control their own future in science (see my previous blog on this).  Indeed, that was a primary message delivered by many of the speakers.  As Dr. Polka said as she started things off, “We are all capable of contributing to change.” 

Check out Joanne's Reddit AMA

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Topics: Career

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 greenfluorescentblog.org 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

Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells

Posted by Kendall Morgan on Sep 30, 2014 4:50:49 PM

Researchers have shown that it is possible to deliver RNA-guided CRISPR/Cas9 nuclease complexes  using adenoviral vectors (AdVs), to a wide range of human cells, including mesenchymal stem cells, and in a rather straightforward manner. These adenoviral CRISPR/Cas9 genome editing tools developed and demonstrated by Manuel Gonçalves and his colleagues at Leiden University Medical Center are now available at Addgene along with a description of their experimental protocol. The three plasmids which have been deposited to Addgene are: pAdSh.PGK.Cas9, pAdSh.U6.gRNAS1, pAdSh.U6.gRNAGFP.

“Although AdVs are being deployed for delivering zinc-finger nucleases into human cells, we think they are still underused in the emerging field of genome editing,” Gonçalves said. “In contrast, AdVs are extensively being explored for genetic vaccination and oncolytic approaches. In genome editing, they are not used much, but we do think they have a very bright future.”

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Topics: Genome Engineering, CRISPR

Sharing Your Lab Protocols: Using Apps to Save Time & Track Your Experiments

Posted by Caroline LaManna on Sep 23, 2014 11:05:27 AM

Recently I learned that Addgene’s pLKO.1 cloning protocol is viewed around 3,000 times each month. I looked this up after trying out the new protocols.io beta platform for sharing, annotating, and storing life science protocols. Since we began sharing this protocol on the Addgene website in December 2006, the pLKO.1-TRC cloning vector (deposited by David Root of the Broad Institute) has consistently been one of the repository’s most frequently requested plasmids. I wondered how many scientists have used the protocol for this plasmid, and if it still being used. After learning of it's continued popularity, I decided this protocol would be a worthy first contribution to the protocols.io community.

Did I mention that this protocol is being used around the world? In the last month it’s been viewed by users in the US, China, India, Canada, the UK, South Korea, and beyond!

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Topics: Scientific Sharing

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