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Special Delivery: Fluorophore Targeting for FRET Studies

Posted by Guest Blogger on Jul 19, 2016 10:30:00 AM

This post was contributed by guest blogger James D. Fessenden, an Assistant Professor at Brigham and Women’s Hospital.

Biochemists often struggle to understand how a protein of interest actually behaves. How large is it? What parts of it move when you feed it substrate or add an essential cofactor? How many binding partners does it have and how do they come off and on in a cellular environment? If these are pressing issues in your laboratory, then FRET experiments are a viable biophysical path to answers.

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Topics: Lab Tips, Fluorescent Proteins

Cpf1 Update: Comparison to Cas9 and NgAgo

Posted by Mary Gearing on Jul 14, 2016 10:30:00 AM

In 2015, Feng Zhang’s lab characterized two Cpf1 nucleases, distant cousins of well-known Cas9. Cpf1 cleaves DNA in a staggered pattern and requires only one RNA rather than the two (tracrRNA and crRNA) needed by Cas9 for cleavage. Now, two new studies show that Cpf1 displays lower off-target editing than Cas9, confirming that this protein is well suited for genome editing. 

Find Cpf1 Plasmids at Addgene

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Topics: Genome Engineering, CRISPR

Generating Mouse Models Using CRISPR/Cas9

Posted by Guest Blogger on Jul 12, 2016 10:30:00 AM

This post was contributed by guest bloggers, Wenning Qin and Haoyi Wang.

CRISPR/Cas9 is revolutionizing the mouse gene-targeting field. Mice have long been extremely useful in the lab – they are relatively small and easy to work with, making them the go-to choice for studying mammalian biology. Similar to any model, mice are not without their problems, but much genetic and physiological data have been accumulated over the years using them. Indeed, the future of mouse work is bright as it is now easier than ever to manipulate the mouse genome using CRISPR/Cas9.

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Topics: Genome Engineering, CRISPR

Addgene at the ASM Microbe 2016 Conference

Posted by Jessica Welch on Jul 7, 2016 10:30:00 AM

We’ve recently begun expanding our presence in the microbiology community. For our first concrete steps into this field, we’ve curated microbiology plasmids from the repository onto one handy Microbiology Resource page and, just a few weeks ago, we attended the American Society for Microbiology's annual meeting (ASM Microbe 2016) for the first time. Our goals at the meeting were to network with scientists in this diverse and exciting field and to find out how we can serve them better. Here’s a little bit of what we learned.

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Topics: Inside Addgene, Microbiology

Plasmids 101: Methylation and Restriction Enzymes

Posted by Marcy Patrick on Jun 30, 2016 10:30:00 AM

Have you ever tried digesting with XbaI or ClaI restriction enzymes and gotten unusual or unexpected results? Or considered why DpnI will degrade your template DNA from a PCR reaction but not the newly synthesized product from a site-directed mutagenesis experiment? The answer to both questions is the same--methylation! Read on to learn about how DNA methylation may affect your restriction digests.

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Topics: Lab Tips, Plasmids 101, Plasmid Cloning

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