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FPbase: A new community-editable fluorescent protein database

Posted by Guest Blogger on May 16, 2018 9:00:00 AM

This post was contributed by guest blogger Talley Lambert, a Research Associate at Harvard Medical School.

The need for a community fluorescent protein database

As recognized by the 2008 Nobel Prize, fluorescent proteins (FPs) have become one of the most indispensable tools in modern biological research.  Any microscopist will tell you that selection of a fluorescent probe (be it an organic dye or FP) is one of the most important steps in the design of an imaging experiment.  The choice is non-trivial, however, as FPs are tremendously complicated entities with a large range of characteristics (color, brightness, photostability, maturation, oligomerization), many of which are dramatically affected by environmental conditions (such as temperature, pH, fusion protein, etc...).  There are many online guides – including an excellent series of posts by Joachim Goedhart on the Addgene blog – outlining various important considerations when choosing a FP, but much of the primary data one might require when making such a decision remains spread across literature in publications that introduce these tools.

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Topics: Scientific Sharing, Fluorescent Proteins

Analyzing CRISPR Editing Results with ICE from Synthego

Posted by Guest Blogger on May 8, 2018 9:00:20 AM

This article was contributed by Jessica Roginsky, Scientific Support Lead at Synthego. Article source: Step-by-Step Guide for Analyzing CRISPR Editing Results with ICE on Synthego’s blog.

CRISPR-based genome engineering has revolutionized the gene editing field by making experimental workflows considerably easier, faster, and more efficient than previous methods. Still, generating reliable results from CRISPR edit data requires the help of robust software tools. As a consequence, a critical step in the gene editing workflow - analyzing the data - is often under-appreciated or over-looked. 

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Topics: CRISPR

Arabidopsis in Education: How the Arabidopsis Biological Resource Center Brings Plant Science to Life

Posted by Guest Blogger on Apr 12, 2018 9:02:19 AM

This post was contributed by guest blogger Courtney Price, the Education & Outreach Specialist for the Arabidopsis Biological Resource Center and the Center for Applied Plant Sciences at The Ohio State University.

Established in 1991, the Arabidopsis Biological Resource Center (ABRC) is one of two global stock centers for Arabidopsis thaliana (Arabidopsis). Our mission is to collect, preserve, reproduce and distribute seeds, DNA and other resources for Arabidopsis and related species. Located at The Ohio State University in Columbus, Ohio, ABRC ships more than 100,000 samples to researchers and educators in 60 countries each year.

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Topics: Plant Biology, Education

When Fidelity Matters: A frank discussion about ligase fidelity

Posted by Guest Blogger on Mar 6, 2018 8:49:43 AM

This post was contributed by guest blogger Greg Lohman, a biochemistry researcher at New England Biolabs.

When do you need a high fidelity ligase—and when is an alternative ligase a better choice? And what is ligase fidelity anyway? Let’s talk about it.

DNA ligases are enzymes that seal breaks in DNA by joining 5 ́-phosphorylated DNA termini to 3 ́-OH DNA termini (1-4). In vitro, ligases (notably T4 DNA ligase) are critical reagents for many molecular biology protocols, including vector-insert joining for recombinant plasmid construction (restriction cloning), adaptor ligation for next generation sequencing (NGS) library construction, and circularization of dsDNA (6). Less commonly utilized in vitro, Taq DNA ligase will ligate only nicks (5-8). Taq ligase is a NAD+-dependent DNA ligase from a thermostable bacterium that can survive high temperatures (up to 95 °C) and is active over a range of elevated temperatures (37–75 °C).

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Topics: Plasmid Cloning

CUT&RUN: An Improved Method for Studying Protein-DNA Interactions

Posted by Guest Blogger on Feb 13, 2018 9:51:55 AM

This post was contributed by guest blogger Matthew J. Niederhuber, a graduate student at UNC Chapel Hill.

Chromatin immunoprecipitation followed by high-throughput sequencing, ChIP-Seq, is the go-to method for mapping where a protein binds genome-wide, and has been widely applied in many model organisms and cell lines. Although ChIP-seq is a relatively simple and robust protocol it does have limitations. The enzyme-based CUT&RUN method overcomes many of these limitations and makes it easier for you to map protein-DNA interaction with limited biological materials.

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Topics: Plasmid Technology, Techniques

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