...expression, but causality and mechanisms remain unclear. Today, targeted epigenetic modification at specific...came epigenome-engineering tools - zinc finger nucleases and TALENs fused to epigenetic modifiers enabled...Christopher Newgard’s Lab has fused LSD1 to the smaller nuclease-dead Staphylococcus aureus Cas9 (Sa-Cas) and deposited...comments! Find Plasmids for Epigenetic Modification   Leah Schwiesow contributed to updating this article. ...

...reducing agent, typically containing thiols, to cleave disulfide bonds. Load the samples, one sample per...conditions from those used by the vendor then it may lead to nonspecific binding.  Whenever possible, include...in making your own knockout line for validation, please view this tutorial from Stuart Orkin's and Daniel...protein causes a thicker band, while less protein leads to a thinner band. It is important to note, however...

...multimer present in your sample. In these instances, please note that the supercoiled dimer band can sometimes... presence of a multimer in your sample remains unclear after visualizing it on a gel, one of the other...consistent with the size of a supercoiled plasmid dimer. Please note that it is common to see other plasmid confirmations...Ausubel FM. Recombination between bacterial plasmids leading to the formation of plasmid multimers. Cell. 1976...

... and the two HindIII digested ends will anneal leaving the 5’ end of your gene just downstream of the ...require you to visualize your DNA using UV light – please note that UV light can damage DNA and that proper... colonies on the backbone + insert plate (or at least more colonies than the backbone alone plate). Unsuccessful...Blog  Additional Resources on the Addgene Blog: Learn How to Verify Your Plasmid Try Out Golden Gate or...

...that introducing a Leu28Cys mutation in eDHFR does lead to covalent binding of the TMP-tag ligand (Gallagher...a different event where CP appears briefly then leaves. Images reused from (Ulrichs et al., 2023) under... if you are comfortable with it or can find a colleague to help you, synthesizing your own ligands can... D., Karassina, N., Zimprich, C., Wood, M. G., Learish, R., Ohana, R. F., Urh, M., Simpson, D., Mendez...

...Want to learn more about genome editing with CRISPRs? Check-out Addgene's CRISPR Resources Learn about...selective cassette minimizes the sequence added, leaving a 34 bp “scar”, but currently requires additional... editing and conditionally manipulate proteins. Learn more at his webpage or follow him on Twitter.   ...

...prevents both the endogenous RecBCD and SbcCD nucleases from digesting linear DNA introduced into the ...E. coli. Exo: Exo is a 5’→3’ dsDNA-dependent exonuclease.  Exo will degrade linear dsDNA starting from...with these promoters (lacI, araC, cI857) to limit leaky expression of the Red system.  Using a plasmid to...the red genes needed for recombineering, but also leads to expression of the int and xis genes which are...

...such as fluorescent microscopy must be used. To learn more about the most suitable fluorescent microscopy...structure of the protein. In some cases this can lead to changes or a complete loss of protein function...more autofluorescence and nonspecific staining leading to false positives. Numerous dyes can be used for...Check out other methods using fluorescent proteins Learn more about antibodies Resources on Addgene.org ...

...epifluorescence microscopy wasn’t able to give a clear view of cellular movement in real time. Two-photon...microscopy has uncovered host-pathogen interactions leading to the understanding of effector function in infected...Weninger. "Dendritic cell behaviour in vivo: lessons learned from intravital two-photon microscopy." Immunology... Get Tips on Choosing Your Fluorescent Protein Learn Some of the Pitfalls of Working With Fluorescent...

... the pPSU2 plasmids are available from Addgene. Learn How to Analyze a Restriction Digest References ...Baby Gel Box Find 5 Smart Phone Apps for Lab Life Learn About the Evolution of Lab Techniques Resources...Kit-Free Alkaline Lysis Plasmid Miniprep Protocol Learn How to Analyze Your Restriction Digests Make Your...

... company, Science with Tom. In this podcast, we learn more about Tom and pick his brain for advice on ...importance in understanding biological history. 4. Please Don't Kill My Hive Beautifully filmed video about...generation science standards and communicate science. Learn more about Tom on his youtube channel and at sciencewithtom.com...

...forward with these efforts, you’ll find new ways to learn about science careers, lab protocols, and, of course...read through each one, plug in some headphones to learn more about the latest and greatest plasmids through...Plasmid webpage. We hope you enjoy this new way to learn about plasmid technologies. If you have any comments...

...journey is at an end! Their name is...(drum roll, please!)... Abi! Thank you so much to everyone who submitted...Which name best fits our new immunoglobulin friend? Please help us decide by voting for your favorite on this...Original Blugene and Aavery would like your attention, please, as they introduce their new friend…  Fig...

...Ready-to-Use CRISPR Lentivirus Learn about Other Members of the Addgene Leadership Team ....     Additional Resources on the Addgene Blog Learn More About Alex's Work with Cas9 Activators Michael...

...Since this is a beta project, it will run for at least 6 months as we assess the needs of the scientific...Browse blog posts from our Plasmids 101 blog series Learn about the different grades of DNA  Read about the...ways to quantify DNA Resources on Addgene.org Learn about our other beta projects Read our Molecular...

...Cambridge to spend some time learning and doing research at Novartis. Listen to learn how Kwabena plans to use...Addgene Blog Check Out Posts on Synthetic Biology Learn about the Addgene, Seeding Labs Plasmid Grant Read...

...to manage these changes quickly and harmlessly. Learn much more about the laws and their possible repercussions...School Advice Part 2: Building Community.” This post leads readers to our latest podcast all about graduate...to catch up on ways to use CRISPR in plants, to learn techniques for detecting genome edits, to discover...

... Now, they’ve filled that gap. As Sakuma and colleague Takashi Yamamoto describe in their recent study...tandem. They went on to show that their CRISPR/Cas9-nuclease and CRISPR/Cas9-nickase vectors work successfully...Currently, the kit only includes two versions of Cas9, nuclease and nickase, but we are now adopting dCas9, dCas9...

...stress-induced biochemical changes that could otherwise lead to depression. And there you have it, mck-PGC-1a1...beneficial effects on the brain,” Ruas said in a press release. “We actually found the opposite: well-trained ... demands,” they write. “In the future, these may lead to treatments for a variety of brain disorders, ...

...corner of this post) is an example of a seemingly clean DNA sequence (no Ns in sight). If you never looked...like these…. 3. Expect to get 500-700 bases of clean reliable DNA sequence Anything less and you might...and small, so I suggest designing your primer at least 50bp upstream of the sequence of interest. 5. Use...