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...TA-based ligation and tagmentation preparation approaches described previously require a preliminary double...that could be used as a template for downstream applications (Berns, 1972). Using qPCR and restriction enzyme... sequencing workflow. Using this simplified approach, we sequenced over 100 AAV vector preparations...believe that this simplified sequencing and analysis approach can be easily incorporated into standard QC regimens...

... “species on species”) immunochemistry (IHC) applications an issue? The culprits are endogenous IgGs in... same species as your primary/sample, and then applying this mixture to your tissue (Tuson, et al., 1990...After washing away unbound fragments, you can then apply your primary and secondary antibodies as normal....Partridge TA (1998). A New Blocking Method for Application of Murine Monoclonal Antibody to Mouse Tissue...

... transfer resources (summaries, electronic MTA approval, archive of old MTAs, and more) Does your TTO ...periodically for possible new awardees, and we’re happy to share stats and strategies with tech transfer...executed between a TTO and Addgene, whose terms apply to all future deposits of materials with Addgene...executed between a TTO and Addgene, whose terms apply to all future materials requests from Addgene MTA...

... unexplored. Using new single-cell sequencing approaches, the lab was able to eliminate the need for axenic...Quake Lab, volunteered to do the sampling and I happily agreed to accompany him to YNP the following year...a thermostable Cas9 be useful? It can enable applications where reagents must be stored at elevated temperatures...stability under harsh conditions. One example application from the publication is the removal of unwanted...

...opportunity to study it and see what happens when technological change happens really fast,” Thompson says. To...of sequence data. A group of scientists from MIT tapped into this data to learn about trends in synthetic...have generated robust tools that have practical applications in monitoring the accidental or intentional ...

...revolution in biological research. We’re incredibly happy to see that one of our career posts has made it ..., we hope we can help all researchers achieve happiness in their careers regardless of their end goals...addgene.org. Thank you again for a fantastic year and happy reading! Top 10 blog posts of 2015 1. CRISPR 101...some common controls and why you might need them. Happy Reading!   Resources on the Addgene Blog Download...

...parasites, to symbionts, to genes and the labels applied to them were vague or contradictory, owing in part...elements. At the time, the use of episome seemed appropriate, especially since the Fertility, or F-factor ...seen as an ubiquitous tool that can be diversely applied to many different experiments. Addgene was founded...Vectors" chapters, and more pictures and pro tips.  Happy reading! Learn more about the history of plasmids...

...Creating a modular, selection-based gene tagging approach Multiple features of C. elegans have made CRISPR...permission from the Genetics Society of America. Applications of this system Dickinson et al. have developed... in cell culture. In addition to CRISPR/Cas9 applications, SEC selection could replace other, more restrictive...Additional Resources on the Addgene Blog An “elegans” Approach to Better Cas9 Editing Efficiency CRISPR 101: ...

...characterize a protein of interest (and we very much appreciate even those small contributions). Of course, no...incorporate FPbase data in their own projects and applications, a REST API allows programmatic retrieval of...within-study photobleaching comparisons.  We very much appreciate feedback, so try out the site and contact us ...Techniques - Which is best for me? A Practical Approach to Choosing the B(right)est Fluorescent Protein...

..., or reordered and PCR enzymes are adjusted. Appropriate fragment and primer design results in covalently...transforma­­­tion or as a PCR template. Some of the applications of the NEBuilder HiFi DNA Assembly Mix include...target gene, for example, are amplified with overlapping regions of homology and inserted into a linearized...associated with otherwise more complicated assembly approaches, and opens the door for new opportunities in ...

...fluorescent signal: untranslated RNAs appear yellow, and translated RNAs appear red. To enable this color switching...and RFP are bound to the reporter, causing it to appear yellow. The act of translation pushes NLS-PCP-GFP...tethers mRNAs to the plasma membrane, it’s not appropriate for use with mRNAs usually translated in a specific...

...millions, if not billions, of dollars, but by approaching it from a new direction, we hope to do it much...Guo, Fengbo Zhou, Paddy O’Leary (UCSF). Our approach is to discover key human proteins that can be ...therapeutics that are safe, effective, and already FDA-approved). This is a big job, but under the leadership ... half the drugs hitting these proteins are FDA-approved or in clinical trials or preclinical studies already...

...in pairing with base editing and prime editing approaches. Recently, Xue Sherry Gao’s lab at Rice University...packaged into viral vectors for potential in vivo applications or multiplex CRISPR screening.   Using tRNAs...and efficient in plasmid and viral vectors. In applications when repetitive tRNA sequences must be avoided...construct DAP array for multiplex gene editing applications, we provide detailed manual and online design...

...research. This year, we put out another call for applications. We had an overwhelming response and were excited...many creative uses of fluorescent proteins. We’re happy to announce the three winners of this year’s Michael...does that mean? Previous research shows that approximately 75% of all protein-coding pathogenic human variants...artificial leaf replica system. “This bottom-up approach will allow me to create a fully controllable and...

...erupted in controversy. Briefly, there are two overlapping, valid critiques being offered of He’s work: ...public involvement, research, and consensus can happen. We just have to look to the United Kingdom’s Nuffield... technique was feasible, three separate things happened nearly simultaneously: The Wellcome Trust funded...biosecurity, and the entire 2018 saga of “what’s happening in genetics this time?” She lives with her partner...

... related to PZA-resistance by using different approaches, including CRISPR and recombination-mediated ...fluorescence microscopy and FACS. In an additional approach, plasmids such as pACBSR-hyg and pFLP-hyg will... the river back to the user.   At Addgene we're happy to be able to support diverse groups of scientists...collaborations. Many thanks to all of the awardees, all who applied, and to Seeding Labs who helped us coordinate and...

.... For a single action potential, GCaMP7s is approximately twice as bright, relative to baseline fluorescence...fluorescence, as the GCaMP7f variant, and is approximately 5-fold brighter than GCaMP6s. jGCaMP7f is the fastest...inactive neurons is reduced. This variant has an approximately 12-fold increase in brightness after ~100 action...genetically-defined neuronal populations with intersectional approaches is a growing field that we aim to support by ...

...engineering E. coli metabolism, maybe you’ll be able to apply your knowledge of metabolism to better understand...favorite FRET-based biosensing technique can be applied to an important neuronal signalling pathway - if...got a useful tool, there’s likely someway you can apply it in neuroscience. AAV tools from Addgene are already...targeting genes to specific cell types in the brain, mapping neuronal networks, and selectively activating signaling...

...spCas9 was the only Cas enzyme widely available and applied by researchers for gene targeting. Fast forward...properties, previously established uses, and intended applications. Within CasPEDIA, there is a classification ...multiplex ability Experimental guidelines and applications Protein structure and properties Links to ...you want to read literature on the less common applications of an enzyme for your non-traditional experiment...

...small, avoidable mistake. Or worse, when you're trapped in a pattern of small errors that significantly...of ways you could change your variables - your approach, your workflow, or your physical setup - to prevent...workflow can have a huge impact. Test different approaches, like whether it's better for you to use cold...resistant to. Just like any other experimental approach, use small, incremental changes to understand ...