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... and after compensation (C) with APC-Cy7.   FMOs FMO stands for fluorescence minus one. It is used...used in the later gates, are more likely to require FMOs. Isotypes Isotype controls are negative controls...daunting process. Controls like single color controls, FMOs, and isotypes can help you more easily gate your...will help you save on sample volume, so you can use most of your samples for flow analysis rather than for...

...signaling molecules. Assay Type Westerns and ELISAs mostly use HRP, a chemiluminescent conjugate that can ...however, is pretty much every other assay you can do. Most non-HRP conjugates are fluorophores, which work ...be detected in the same assay. This is one of the most complex subjects in antibody-based protocols - but... them whenever possible. This will allow for the most accurate read and reduce needed compensation.    ...

...research needs. In this update, we will focus on the most recent advances and how some of these variants may...PAM sequence recognized by the nuclease. For the most frequently used Cas9, spCas9 from Streptococcus ...pX-evoCas9, which they showed also increased specificity. Most recently, the Jungjoon Lee Lab used directed evolution...Recent alternatives to Cas9 While Cas9 remains the most widely used CRISPR nuclease, recent work has shown...

... is the most important resource that Addgene provides the scientific community? Katz: The most important...required to do research. Paratore: What I’ve been most impressed with is the level of proficiency that ...science? What about the future of open science are you most excited about? Katz: What’s exciting about the open...

... envelope expressing plasmid, such as our second most requested plasmid of all time, pMD2.G. Find psPAX2...this plasmid. Indeed the first version is the 15th most popular plasmid of all time, but was found to have...available through Addgene in 2015, making this the most recently available plasmid in the top 15. Find pSpCas9... your gRNA. It’s the former version of our sixth most popular plasmid pSpCas9(BB)-2A-Puro (PX459) V2.0...

... on Addgene.org Addgene's most popular lentiviral plasmids Addgene's most popular retroviral plasmids...discuss them together. Gene expression The reason for most viral experiments is to deliver a genetic cargo,...Fun fact: There are 57 human adenovirus types, but most vectors are based on adenovirus 5 (Ad5).   ...

...sleeves and most patients still don’t benefit from this approach. One central bottleneck is that most recurrently... and most of the individual variants found in tumors are exceedingly rare. As a result, how most of these...

...labs, and are often impractical for small projects. Most researchers instead turn to so-called “mismatch ...(such as droplets or wells on a chip), such that most partitions will contain either one or no copies ...functional knockout” of a gene of interest is one of the most common applications of precision genome editing,...mono-allelic and bi-allelic mutations: Perhaps the most noteworthy advantage of digital PCR assays is that...

... guides around the region to find the most efficient guide (most guides would work, but they sometimes...Scientist, Matt Ferenc, to organize the CRISPR forum's most popular questions and answers. Designing your CRISPR...multiplexing CRISPR to target multiple genome loci, the most efficient and easiest way is to co-transfect several...homology arms be for efficient recombination? A10: The most we've tried to insert so far has been 1 kb. We used...

...take a quick flash back to 2004. “Shrek 2” was the most popular movie in theaters, the Razr was the hottest...request for a kit came on March 9, 2005. It took almost six years to reach our first major repository milestone...distributing ready-to-use viral vectors from some of our most popular lentiviral and AAV vectors, as well as developing...guys rock." — Andrew York, Calico Life Sciences. Most of all, we’re incredibly thankful for all the scientists...

...features would allow for the best performance in the most contexts (Figure 1). Together the features with ... situation (Devaraj & Finn, 2021). In one of the most common Click Chemistry reactions, one component ... But the reactive groups of Click Chemistry are almost completely absent from living systems. What are...the ER and Golgi apparatus and outside the cell, most cysteines are oxidized in disulfide bonds. Reactions...

...good idea. The scientific community is loving it. Almost 10,000 viral vector samples were distributed in...Last time I checked we were ranked as the 21,806th most popular website in the world. I know, right? Upgrades... at Addgene is about half of what is standard in most sectors. The Addgenies support a high collaboration...a 90 person company. We were and remain ahead of most companies in creating a welcoming, inclusive workplace...

...harder to calculate, in both our 2018 and 2023 poll, most people said it took them more than 22 hours, so ...consumables today. Finally, this doesn’t account for the most valuable component in science – the time spent by...your lab (but if you need a lot of plasmids, it’s almost always faster to order from us!) Ask the PI If ...a collective and very cool 8,768,560 hours – or almost exactly 1000 calendar years!   *Dammit, man, I...

...resistance, restriction sites used during cloning, and most importantly, the predicted full plasmid sequence...ready to sequence verify the plasmids. This is the most important step in ensuring quality, as it will catch...particular plasmid you are working with, but, in most cases at Addgene, we check what we call the “insert...Important features may include cloning junctions (where most sequence assembly errors occur), deletions, mutations...

...transgene expression across several plasmids. The most commonly used viral vector systems are made up of...Follow our viral vector blog posts for updates on the most recent deposits, protocols, and tips for using retroviral... Please check www.genome-engineering.org for the most up-to-date information. Check our CRISPR guide and... and blog to learn more about CRISPR and the most recent developments in the genome editing field. Lentiviral...

...in a direct and indirect ELISA.   Sandwich ELISA Most commercially available ELISA kits are for a sandwich...between two antibodies in a sandwich ELISA. While the most common type of competitive ELISA is a modification... ELISA well is based on a colorimetric reaction. Most commonly, this is a blue reaction that results from...Jagarlamudi et al., 2015.   The maximum OD that most microplate readers can detect is 4.0, so if you ...

...also see who an author is most influenced by (other authors who they cite the most), and who the author influences...particular authors, with the authors who published the most papers with your keyword populated at the top of... influences (who cites the author the most). Best uses Finding papers frequently cited by others. This...

...just trying to shred a long non-coding RNA, cutting most anywhere within the RNA will do. Similarly, if you...Kwart et al, 2017). To add insult to injury, the most utilized PAM sequence, that of SpCas9, is NGG. Assuming...Have we reached an essentially PAM-free solution? Almost! Assuming a PAM of NRN, 50% of randomly selected... or take a bp or two upstream or downstream) for most gene targeting experiments. Between SpRY and several...

...potential for correcting monogenetic diseases. Now, almost 10 years later, CRISPR’s potential for treatment...thalassemia (TBT), both of which affect hemoglobin. Most of the targeted treatment strategies for these disorders...shared from these trials has been outstanding, with almost all patients experiencing sustained and substantial...hATTR patient outcomes. Bacterial infection and HIV Most CRISPR therapies have been targeted to precisely...

...Calcium is arguably one of the most important metal ions in biology (sodium and potassium are up there...regulated. In this blog we will explore some of the most commonly used as well as newly developed calcium...dramatic. You want to use a calcium sensor that is most sensitive to your expected range. For example, calcium... often delivered using viral vectors! One of the most significant advantages of viral delivery is the ...