...approaches everything; we asked ourselves, “How can we do this in a way that most helps scientists and allows... the service and figure out what we can afford to do and when. Addgene’s Legal Team acquired permission...impression that it took a lot of work and dedication to do this well ... because it did. So finally, a big thank...

...enable efficient base editing and prime editing. How do you then set up an MBE system that contains a double-win...editing, while keeping the system to minimal size. To do so, they harnessed the tRNA processing mechanism,...exhibited reduced Cas9-dependent off-target editing and do not cause higher Cas9-independent off-target editing...

...vector propagation and sequencing a headache. How do I maintain ITR integrity and prevent unwanted mutations...reduce the likelihood of ITR deletions occurring.  How do I validate my ITR sequences?  Next-generation sequencing...immediately outside of the ITRs in the plasmid backbone and do not properly assemble, but these sequences can sometimes...

...worthwhile. Particularly if in the end all they want to do is to quickly generate a single cell line and proceed... Advisory Board) says: "When you're setting up to do a genetic screen, you need to focus on building a...assay is scalable and robust. What you don't need to do is spend your time troubleshooting reagent production...

...in the article are those of the guest blogger and do not necessarily represent the views of Addgene. I... 3. NEB Tools by New England BioLabs In my lab we do a lot of genetic construction and cloning experiments...learn about biochemistry. The least this app will do is allow you to write your colleagues’ names in a...

...introduces himself and explains why he decided to do an MD PhD - it's all in the family. 3:56 - 7:04 :...gene drives work and what work he's planning on doing with CRISPR-based gene drives focusing on safety...

...Duedu inspire new Ghanaian scientists to continue to do excellent work in the biological sciences. Future...plasmids from the Addgene community and hope we can do even more to enable scientists in developing regions...

...they do in the software. Most sgRNA design tools require the user to specify an organism. They do this...what the parameters represent. The best learn-by-doing tool is E-CRISP. The authors have made a huge effort...for labs: Define what you need the software tool to do before going shopping for one. Use the CRISPR Software...criteria. Repeat until you have found the best tool. Do several experiments and use the results to re-evaluate...make a difference in our experiments and lives. To do this effectively, there needs to be good communication...

... cost. The problem with these assays is that they do not provide sequence-level data. They also have a... a limit of detection of ~5%. This means they do not reliably detect editing events that occur in less...variable) Sequence-level data, extremely sensitive Do not sequence all DSBs, may miss unpredicted off-target...Tsai et al. 2015). Additionally, current algorithms do not take into account other elements, including those...GUIDE-seq and BLESS detect double-stranded breaks and do not require high sequencing read counts making them...

...take to reduce them. But we do know we will continue to ask “What can we do to make our materials both ...by researchers all over the world — and we really do mean all over (Figure 3). We continue to add new ...more reproducible science and learn what Addgene can do to help. More resources on the Addgene blog Twenty...