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Plasmids 101: Screening Strategies Used in Plasmid Cloning

Posted by Jennifer Tsang on Sep 15, 2020 9:15:00 AM

If you’re cloning a plasmid, you’ll need a way to find the needle in the haystack: the one perfect clone that contains the plasmid you’re looking for out of the many cells that don’t. One way to begin the search is by using selection strategies, where only cells that have gained or lost a specific gene survive (ex: antibiotic resistance marker). In a previous blog post, we covered how to use positive and negative selection in plasmid cloning

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Topics: Plasmids 101, Plasmid Cloning, Plasmids

Addgene’s Newest Way to Speed Your Research: The Cloning Grade DNA Service

Posted by Paolo Colombi on Jul 28, 2020 9:15:00 AM

What if after ordering a plasmid you didn't have to grow the bacteria and prep the plasmid before you begin your cloning experiment? What if after receiving the plasmid from Addgene you could directly start your digest, PCR, or transformation? 

Great news. A subset of our collection has been prepared as part of our new cloning grade DNA service and is ready to use out of the box.

We estimate that providing cloning grade DNA could save scientists up to three days of work. We’ve taken care of all the steps going from streaking to a single colony to DNA extraction and quantification so that you can focus on what you want to do with your plasmid without spending time and resources in preparative experiments. 

We’re excited to begin offering this service to further contribute to our mission to “accelerate research and discovery.”

Here are the details.

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Topics: Addgene News, Plasmid Cloning, Plasmids

Plasmid Preps: Different Purity, Different Quantities, Different Uses

Posted by Paolo Colombi on Jun 16, 2020 9:25:46 AM

Not all plasmid preps are the same. Before purifying a plasmid from a bacterial culture, it is important to consider your experiment.  It will dictate the amount of DNA you need, and at which level of purity. Based on these premises we can classify a plasmid preparation in 3 different ways:

  • Transformation grade DNA
  • Cloning grade DNA
  • Transfection grade DNA
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Topics: Plasmid Cloning, Plasmids

Early Career Researcher Toolbox: Free Online Molecular Biology Tools

Posted by Beth Kenkel on Apr 14, 2020 9:15:00 AM

Primer design. Plasmid mapping. DNA sequence analysis. We all have our favorite tools for tackling these particular tasks, but they tend to be scattered about the internet. To help you keep your virtual molecular biology toolbox organized, today’s post features a list of free online molecular biology tools all in one place.

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Topics: Plasmid Cloning, Plasmids, Early Career Researcher

Troubleshooting Your Plasmid Cloning Experiment

Posted by Guest Blogger on Sep 24, 2019 8:55:00 AM

This post was contributed by Oskar Laur, head of the custom cloning core at Emory University, and Paolo Colombi, a product development scientist at Addgene.

Cloning can be quite an arduous process. The PCR could fail to produce a product, the transformation may not result in any cells, or all the colonies screened might not contain the correct plasmid. There’s a lot that can go wrong! With all the steps in the cloning process, there are also many ways to troubleshoot the cloning experiment. Here are some tips that will help you with your cloning project, and hopefully obtain your coveted plasmid with no substantial delays.

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Topics: Plasmid Cloning, Plasmids

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