...mediated knock-ins in mammalian cells. J Biotechnol., 16(4): 387-390 (2020). 10.1016/j.jbiotec.2019.11.010 Wright...CRISPR Featured Topic Page Learn about Non-Homologous End Joining Learn about Base Editing Read 3 Tips to ... post on increasing HDR efficiency to read more about optimizing this distance. Homology arms and types...screening strategies for your experiment. Also learn about Addgene’s knock-in plasmids for reagents to help...

...the same sample, you could end up with a titer of 4 x 1012 genome copies/mL or 8 x 1012 genome copies/... droplet generator, you will have to be careful about transferring your droplets from the droplet generator...Additional resources on the Addgene blog Read more about polymerase chain reaction Browse our blog posts ...collection of viral vector related protocols Learn more about the Addgene viral service ...

... PMID 25417800. PubMed Central: PMCID PMC4282009. 4. Yosef, Ido, Miriam Manor, Ruth Kiro, and Udi Qimron...organizations are working to help educate the public about proper antibiotic usage. Although these steps are...laudable, they likely won’t solve the growing problem. Above all, we need new antimicrobials, ideally ones for...used to deliver any of the three systems discussed above, with the caveat that phages are very diverse, and...

...endotoxin-free plasmid purification kits.     Figure 4: Schematic of the barcode matching system and PCR-... before being distributed to customers. Details about our production protocols, titering methods, and ...either in-house by Addgene scientists or through collaboration with viral vector manufacturing facilities, ...generated from lentiviral preparations (described above) that are subject to a concentration step prior ...

...beads are collected by centrifugation or a magnet. 4) Unbound proteins are washed away. 5) The target protein...proteins or nucleic acids. Read on to learn more about this versatile technique.  Immunoprecipitation ...proteins in an IP are not denatured (learn more about antibodies here!) When using Protein A/G to immobilize...eluted by boiling the beads in SDS. (To learn more about Western blotting check out our blog post The Basics...

...dissolved in DMSO which allows both biotin and biotin-4-fluorescein to be dissolved at a higher concentration... plasmids you need. If you'd ever like to write about a recent plasmid deposit please sign up here.  Here's... a cell. The Goedhart lab, Gadella lab, and collaborators have an ongoing effort to improve genetically..., that are optimized for excitation wavelengths above 1,000 nm and enable two-color imaging with red fluorescence...

...Nature Genetics 28, 92-95. PubMed PMID: 11326284. 4. Bennett J, 2017. Taking Stock of Retinal Gene Therapy...famously visited Congress to help increase awareness about the potential of gene therapy!). It would take another...muscular atrophy, neurodegenerative diseases and metabolic diseases are also underway and reported results...

...PMID: 18355003. PubMed Central PMCID: PMC2802578. 4. Xue, Lin, et al. "Imaging and manipulating proteins... the frontiers of biological imaging.  *Worried about your budget? Email [email protected] Resources on the Addgene Blog Learn About Other Protein Tags Catch Up on Your Fluorescent ...

... aversion/anxiety. The mouse can move freely on a 4-arm cross and choose to stay on an open arm (risky...distant regions. They also don’t give any clues about the types of neurons involved or whether their involvement... Viral Vectors Resources on Addgene.org Learn about Addgene's viral service Find viral protocols and...

...PMC5526198. Find plasmids from this paper at Addgene. 4. East-Seletsky, Alexandra, et al. "RNA Targeting by...potential applications of Cas13a given what we know about its structure and function? For starters, Cas13a... Check Out Our CRISPR Featured Topic Page Learn about Anti-CRISPRs Multiplex Genome Editing with Cpf1 ...

...276.32 (2001): 29621-29624. PubMed PMID: 11408473. 4. Subach, Fedor V., et al. "Monomeric fluorescent timers...Additional Resources on the Addgene Blog Learn More About Fluorescent Protein Aggregation Catch Up On Your...

...Mauch,  K. Deininger, et al. (2007). Nature Methods, 4, 331–336. Pubmed. P. Keller, M.B. Ahrens.  (2015) ...protein (GFP) (6).   The Dodt and Frank Bradke laboratories later developed the 3DISCO (3-dimensional imaging...

...histories, accurate to within 40 minutes. Figure 4: Time-resolved protein ticker tape. A) Design of iPAK4...section of the Addgene antibody page, and learn more about the whole-brain reference atlas efforts, including...

...transcriptional activation screening. Nat Protoc. 2017 Apr;12(4):828-863. doi: 10.1038/nprot.2017.016. Epub 2017 ...ordering a pre-made library, you don’t need to worry about amplifying and producing lentivirus. But pre-made...

...just need to figure out how to make it suspenseful. 4. Start preparing your talk early - like today Don’...interesting content Talk about something you know a lot about and that you care about. Don’t try to present...engaging and memorable. If you read my story above about the presentation faux pas, you are more likely...the room, so it is a good idea to start thinking about public speaking early and often. Two of my jobs ...dynamite presentation?  To that end, let’s chat about giving science talks and how to make them serve ...other people give great talks We are not talking about lab meeting here. Lifeless presentation of experimental...You want to give the audience something to think about, argue with, or learn from. Some of the best talks...

...general mission to share- as opposed to sell- science. 4. You can prepare answers beforehand. Joanne answered...not too worried about selling yourself and really do just want to get information about yourself and your...where users can participate in forum discussions about a wide variety of topics. Individual forums are ...where users can ask designated scientists anything about a particular topic. Given Joanne’s expertise in... is well moderated. One of our original worries about sharing science advice on Reddit was that we’d be...answering questions. r/Science has pretty strict rules about not being too sales-y in posts which falls in line...any scientists to refer to if they have questions about science careers, women in STEM, work-life negotiation...

...Science Talk." 4. The chalk talk, simple in principle The first question I usually get about chalk talks ... potential colleagues study. Think about how you might collaborate or, at the absolute minimum, can appreciate...the first sentence or two what your research is about and what you think you can accomplish. No matter...your seminar accessible. Let the experts ask you about the details and then you can display your deep knowledge...expert's question than to leave everyone confused about your research because you waded too deep into the...obstacles, alternative approaches, potential collaborations, critiques of your postdoctoral research, etc...effectively convey your message. Key Things to Think about When Developing your Chalk Talk These items should...

...Plasmid incompatibility. Microbiology Reviews. 1987;51(4):381–395. Pubmed PMID: 3325793 Schumacher MA (2012...101 blog posts Learn more about the origin of replication Find blog posts about plasmid cloning Resources... plasmids within the same bacterial cell, think about whether plasmid incompatibility will affect your...this is expanding as we obtain more genomic data about plasmids (Shintani et al., 2015). Now, bioinformatics...of the microbiological world. (Want to know more about these plasmid types? Read our Plasmids 101: Environmental...

...extremes lies the linear detection range. Figure 4: The linear detection range of the assay is between... this chain (blue) to infer or calculate things about earlier ones. In practice, multiple sources of error...condition 2.” By eye, the loading control appears about the same between the two conditions. But it isn’.... However, it throws both assumptions discussed above into question. Assumption #1: The loading control...searching the literature to see what is already known about your housekeeping protein in your context. If you...step from the POI blot, you don’t need to worry about the linear detection ranges of multiple targets....

...null clone, you should screen, at the very least, 4 clones. Did you use a dual guide system to generate... your edit, it’s important to know a few things about your target cells and the desired edits. First, ...et al., NAR). It works just like TIDE (outlined above), except that it uses a 3rd sequencing trace file...great option, especially if the options outlined above aren’t realistic. Check Out Our Post on Sequencing...higher probability locations. If you’re worried about off-target effects, you can consider using a version...SpCas9-HF1, eSpCas9(1.1), or HypaCas9. Read more about Cas9 options in our blog post here. Dealing with...