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...viral genome sequencing (VGS). Why should you care about viral quality control? AAV vectors are produced ... let us know if you have any specific questions about viral DNA NGS you’d like us to discuss. We are also...or at [email protected] to answer your questions about our quality control process! The images in this...

...inserted GFP is expressed (knocked in). As seen in the above example, you can remove a floxed resistance gene...Get the Basics of Homology Directed Repair Read about Generating Knockouts with CRISPR through Non-Homologous...Expression Vectors Find Cre-Lox Plasmids Learn More about CRISPR  ...

...sold by Clontech and is in a freezer box in just about every lab in the world, is not inert. In all seriousness....11 mM [2]. Following the simple logic outlined above, you might think EGFP is unlikely to form dimers...affinity assays used in the papers describing the FPs above including molecular sizing columns and native gels...

...preparation from Addgene, you can skip the steps above! Cells are then transduced with lentivirus containing...practices, CRISPR libraries can help you learn a lot about your phenotype of interest. For more detailed information...Libraries” (McDade et al., 2016). To learn a bit about how CRISPR/Cas9 can be used in other types of experiments...

...Korea, and beyond!   Check Out How Addgene Thinks About Developing Lab Management Software    Organizing...of other scientists using pLKO.1 variants learn about those updates? This is a problem many scientists...our Plasmids 101 blog series. Want to learn more about sharing your plasmids via Addgene? Check out all...

...such as full plasmid sequences provided by the laboratories that deposit their plasmids with us or other...mismatches and gaps in the alignment. If you are curious about the differences in the blastn programs, check out...Inside Look at NGS Plasmid Quality Control Learn about our Snapgene-powered plasmid maps. Resources on...

...by replicate values provides useful information about the accuracy of the curve, which we will discuss...many scientific applications users aim for an R2 above 0.95.    Additional considerations When running ... or make smaller dilutions.        As mentioned above, it is a good idea to run the standard dilution ...

...Blugene (check back on December 5 to learn more about this!). I’ve assembled a series of screenshots ...search. These pages provide helpful information about the subject, link to related guides and protocols...Additional resources on the Addgene blog Learn more about other features on Addgene's website ...

...CRISPR Featured Topic Page Learn about RNA Targeting with Cas13a Learn about CRISPR DNA Base Editing Resources...and sequences that can be edited.   As discussed above, the ability to edit RNA has multiple advantages...

... 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 ...

... J. (2002) Arabidopsis : a laboratory manual. Cold Spring Harbor Laboratory Press. Available at: https...https://books.google.at/books/about/Arabidopsis.html?id=IfZAMNPWVk4C&redir_esc=y (Accessed: 21 September ...

...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...

...Plasmids 101 Learn about genome-wide screening with CRISPR and Lentivirus Read about the ClonTracr Library... the proper cell line is used; several common laboratory cell lines including 293 were immortalized with...

...more feasible, the lab of Alessio Ciulli and collaborators developed CRBNmidi (Kroupova et al., 2024). ...stepwise, achieving mutation efficiencies of up to about 9% of colonies (102 to 104 times higher than traditional...Bentley Time to get flexible! Alex Dunn’s lab and collaborators have deposited a new mechanosensor called PILATeS...

... negative regulators. The genome of E. coli is about 4.6 million base pairs long and contains a single...copy number and compensating for the additional metabolic cost imposed on the host with a beneficial function.... On the other hand, they also impose a lower metabolic cost on cells and produce lower amounts of protein...

...Generating a transgenic plant takes even longer, is more labor intensive, inefficient, and often not reproducible... region is transferred into the plant. In the laboratory we can utilize this natural process by replacing...system. The vectors mentioned in this blog post above are ready to use vectors, but with the MoClo system...

...usage bias Read more about plasmids in our Plasmids 101 blog series Learn more about trends in synthetic... out Addgene's Molecular Biology Reference Read about genetic code expansion Browse our plasmid collection...

...Addgene Blog Learn more about Cas9 Variants and the Expanded CRISPR Toolbox Read about CRISPR enzyme Cpf1 ... and specific? The two AAV strategies described above showed successful target modification, indicating...

... 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...

...In our last post, we talked about the first base transversion editors: CGBEs, or C → G Base Editors. ...apyrimidinic sites and abbreviated as AP sites. A collaborative team from the David Liu and Dali Li labs reported... CRISPR 101: Cytosine Transversion Editors Read about four base editor reporters Resources on Addgene.org...

...much information as possible from depositing laboratories and associated publications, but our quality...the process. Another design choice we appreciated about Nextflow was how effortless it was for us to switch...interface. In our next post, we’ll share our story about how we used Nextflow Tower to augment our pipeline...

...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...

...series of one- or two-marker plots, like in Figure 3 above, until the population of interest has been successfully...  Of course, there are many more things to learn about when diving into flow cytometry data. There’s different... ways to use FSC and SSC, as briefly alluded to above. There’s many variations in how one can gate, select...

... 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 ...

...successfully to derive iPSCs from PBMCs. Most laboratories use Sendai virus to reprogram PBMCs efficiently...establishment of virus-free hiPSCs usually require about 5-10 passages. Furthermore, working with Sendai ...source of somatic cells. This translates to only about 16 hiPSC colonies per 100 ml of starting blood sample...

...or postdoc forever, and you need to be thinking about what comes next. To that end, one of the most important...personal connection; the importance of which will be elaborated on later. As for what job fairs to consider, ...the specific position he or she originally called about, you can keep in touch and reach back out when you...

... more about CcdB, the toxic key to efficient cloning Read all Plasmids 101 blog posts Read about knockout...with the other toxin-antitoxin systems described above, one part of the pair is more stable than the other...

...following unanswered questions about CRISPR screen design. Unanswered questions about genome-wide CRISPR/Cas9...Morgens et al drew some generalizable conclusions about off-target activity, demonstrate safe-targeting ...

...enable accurate labeling of neurons. As discussed above, weakly expressed proteins in particular are difficult...resembles images of the deity (see comparison image above). The church of the Flying Spaghetti Monster is ...tags. 4. High-resolutions microscopy As indicated above, smFPs can be used with conventional microscopes...

... on the Addgene Blog Learn More about FLEX-vectors Learn More about Cre-Lox Check out Retrograde AAVs...expression have been developed. Read on to learn more about FLP-FRT, Cre-lox, and how combinations of FLP and...