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Hot Plasmids April 2018 - Protein Degradation, Nanoscopy, FIRE-Cas9, and Yeast Expression Tools

Posted by Various Addgenies on Apr 23, 2018 10:00:31 AM

Every quarter 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 they'll make it easier for you to find and use the plasmids you need. Below you'll find our hot plasmid articles from the first quarter of 2018. If you'd ever like to write about a recent plasmid deposit please sign up here.

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TRIM-Away: Targeted Endogenous Protein Degradation

Article contributed by Mary Gearing

CRISPR and RNAi have helped researchers alter DNA sequence and RNA expression - but what if you just want to change protein levels? Until recently, no such tool existed for rapid, customizable protein degradation. Enter TRIM-Away - an antibody-based system developed by the James and Schuh labs. Clift et al. applied TRIM-away to nine proteins in ten ddifferent cell types, including difficult-to-manipulate primary cells, achieving knockdown within 1-2 hr of reagent delivery. TRIM-Away is suitable for both short-and long-lived proteins, and it can even distinguish between protein variants when used with a variant-specific antibody.

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Topics: Hot Plasmids

Plasmid Cloning by PCR

Posted by Various Addgenies on Mar 29, 2016 10:30:00 AM

In its simplest form, PCR based cloning is about making a copy of a piece of DNA and at the same time adding restriction sites to the ends of that piece of DNA so that it can be easily cloned into a plasmid of interest. You can use similar processes to add overhangs to your insert of interest for Gibson assembly. The steps following primer design and the PCR process itself are very similar to those outlined in our restriction cloning post with a few quirks specific to the PCR cloning process - please check out that post if you need a more detailed refresher on the downstream steps.

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Topics: Protocols, Plasmid Cloning

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