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Another Pathway into Cells: iTOP

Posted by Mary Gearing on Jun 23, 2015 4:37:00 PM

Primary cells recapitulate the natural biology of a cell type of interest better than immortalized lines derived from the same cell type; however, their usage has been limited by technical problems. For instance, it’s much more difficult to introduce a gene of interest into primary cells, so most primary cell lines require viral infection. A new paper from Niels Geijsen’s lab suggests that primary cells may be better transduced using only protein. Read on for a description of the lab’s iTOP protein-only transduction method and its potential applications to CRISPR/Cas9 genome editing.

 

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Topics: CRISPR, Techniques

Easing the Protein Purification Process with pCri

Posted by Mary Gearing on Jun 19, 2015 11:08:00 AM

Protein purification can be one of the most stressful lab activities. Working with proteins requires a substantial amount of properly folded, relatively pure protein, but getting to this stage is often much easier said than done. As reviewed in our Plasmids 101 series, proteins are overexpressed from a plasmid construct, most often in special E. coli strains designed for protein expression. Cultures are then lysed and the protein of interest is purified using an affinity tag. Additional tags may be used to improve protein stability and solubility.

Determining the best way to express one’s protein of interest can save a lot of time later.

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Topics: Hot Plasmids, Plasmid Kits

Science Communication Snapshot: DayCon 2015

Posted by Mary Gearing on Jun 17, 2015 5:04:11 PM


SITN DayCon 2015 Team

Here at Addgene, we’re dedicated to advancing and sharing science! In association with the Harvard graduate student organization Science in the News (SITN), we recently sponsored a first-time event called DayCon. DayCon is a one-day conference aimed at the general public that provides an accessible introduction to various scientific topics. Over twenty graduate student volunteers worked hard to make this Saturday event a success, a true testament to the commitment of SITN members.

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Topics: Scientific Sharing, Science Communication

More Data for You: Find Articles Citing Addgene Plasmids

Posted by Caroline LaManna on Jun 16, 2015 10:15:40 AM

Exciting news! Addgene recently rolled out a new feature on our plasmid pages - links to articles citing this plasmid. Now you can learn how a plasmid has been used by multiple labs and see what experimental systems it has been validated in. 

If a plasmid's Addgene ID # has been referenced in other publications, you'll find a link to the list of citing articles under the "Resource Information" heading in the right column of the plasmid page. Check out the purple arrow in the screenshot below to see what I mean.

Additional Features

Once you've clicked on the "# References" link under the "Resource Information" heading, you'll be directed to a page listing the articles that cite this plasmid. You can use the dropdown to increase the length of the list (purple oval in the screenshot below). You can also use the "Search Table" box at the upper right of the table to search and filter the list of citing articles. From the article list you can click on the PubMed link to find the article abstract and more.

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Topics: Hot Plasmids, Scientific Sharing, Inside Addgene, Using Addgene's Website

Plasmids 101: Blue-white Screening

Posted by Jessica Welch on Jun 4, 2015 9:03:00 AM

This post is part of our ongoing Plasmids 101 series. Plasmids 101 will provide you with an overview of general molecular biology knowledge and techniques. If you are interested in reading more, you can find the rest of the Plasmids 101 posts here.

Now that we have covered antibiotic selection here at Plasmids 101, we can talk about an even more specific method of screening your cloning reaction. Being able to select for colonies that contain your plasmid is a great start when cloning, but how about being able to choose those that contain plasmid with an insert? Blue-white selection is a widely used method to do just that!

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Topics: Plasmid Elements, Plasmids 101

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