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What's the Best Way to Elute and Store Your Plasmid DNA?

Posted by Paolo Colombi on Jun 4, 2020 9:22:22 AM

If you use a kit for DNA purification or if you use a DIY purification protocol, you might have noticed that there are many options to elute your DNA prep. You might see protocols that recommend eluting in water, Tris-EDTA (TE), just Tris buffer, or some other variations. Does it make a difference? Short answer: yes. This last step will influence the stability of your sample over time and determine which experiments you can effectively use the DNA.

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Topics: Molecular Biology Protocols and Tips, Plasmids

Hot Plasmids - June 2020 - Barcoded CRISPR Library, Sparse Cell Labeling, Calcineurin Reporter, and DNA Staining Dye Alternative

Posted by Various Addgenies on Jun 2, 2020 9:15:00 AM

Every few months we highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles provide brief summaries of recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need. If you'd ever like to write about a recent plasmid deposit please sign up here

 

 

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Topics: Hot Plasmids, Other Plasmid Tools, Plasmids

New and Upcoming Viral Vectors - May 2020

Posted by Maya Peters Kostman on May 28, 2020 9:15:00 AM

Since the beginning of our viral service in 2016, we’ve added many new tools to our inventory of ready-to-use viral vectors. Here are some of the AAV and lentiviral preps we have released in the last few months:

  • Lentiviral preps for expressing SARS-CoV-2 proteins
  • ChR2 for targeted expression
  • DREADDs
  • GFP dependent Cre and FLPo
  • And more! 
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Topics: Viral Vectors, Addgene’s Viral Service

Fluorescent CRISPR Reporters: SRIRACCHA and GEmCherry2

Posted by Alyssa Cecchetelli on May 26, 2020 9:15:00 AM

If you’re using CRISPR to make a genome edit, how do you know if your CRISPR experiment was successful in your organism or cell type? You can use DNA sequencing or other molecular cloning techniques to determine CRISPR/sgRNA efficiency of an experiment and confirm the correct edit was made to the genome without any off target effects. However, these methods can be labor intensive and quite time consuming.  

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Topics: CRISPR, Other CRISPR Tools

Genetically-encoded Sparse Cell Labeling - A SPARC of Innovation

Posted by Aliyah Weinstein on May 21, 2020 9:15:00 AM

Until recently, there were no completely genetic tools that would allow researchers to label just a fraction of a single genetically-defined subset of cells. By labeling fewer cells in a population, it’s easier to visualize individual/non-overlapping cells. While transgenic animals are commonly used to specifically manipulate a cell type of interest in an organism, all cells of that type are affected. Previous tools to overcome this restraint include an AAV-based sparse labeling system where a limiting amount of AAV are injected into the brain of Cre-expressing mice to trigger recombination and expression of a transgene (Lin et al., 2018). However, this system requires precise titration of the AAV. So far, tools to overcome this challenge in Drosophila require heat-shocking the system or using chemical inducers of gene expression (del Valle Rodríguez et al., 2011). 

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Topics: Fluorescent Proteins, Fluorescent Imaging

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