...mechanical work for editing. Class 1 CRISPR systems make up the majority of CRISPR-based immune systems in...broken down into different types, with Cas3 systems making up type I, the most abundant type in the Class ...away at both the non-target and target strands by making cuts every so often. Cutting both strands ensures... for Cas3’s biggest trick. Cas3 likes to bluff, making you into think it’s more powerful than it is. The...issues with expressing all Cascade components can make Cas9 easier to work with than Cas3. Future targets...1477. https://doi.org/10.1038/s41587-019-0310-0 Makarova, K. S., Wolf, Y. I., Alkhnbashi, O. S., Costa,...

... we’d like to make sure they are present at a very low concentration. So how do we make sure to catch ...an experiment because of poor reagent quality. To make sure our AAV vectors are of the highest quality,... mission of accelerating scientific research. To make sure that adeno-associated viral (AAV) vectors available...robots. Unfortunately, humans have been known to make mistakes. What if a tube is mislabeled or someone...K, Sanders E, Tasissa M, Kostman M, Tillgren M, Makana Hanley L, Mueller I, Mitsopoulos A, Fan M (2020...

...be “let’s try to make this STEM activity fun,” but rather “let’s see how we can make this fun activity...time is giving fewer examples and think of a way to make this section more interactive. If you can think ...good analogy that involves student participation, make it happen! How should we use CRISPR? How shouldn...students that scientific history was very much in the making. One of the exciting, and challenging, aspects ... Perhaps you have young children and you want to make sure their STEM education is supplemented. A 2009...

...Zetsche B, Gootenberg JS, Abudayyeh OO, Slaymaker IM, Makarova KS, Essletzbichler P, Volz SE, Joung J,...Non-SpCas9's bind a variety of PAM sequences, which makes them useful when no suitable SpCas9 PAM sequence...non-SpCas9's may have other characteristics that make them more useful than SpCas9. For example, Cas9 .... BV3L6 Cas12a, AsCpf1, uses a 5'-TTN PAM which makes it easier to target AT-rich genomes (Kleinstiver...Gootenberg JS, Kriz AJ, Zetsche B, Shalem O, Wu X, Makarova KS, Koonin EV, Sharp PA, Zhang F (2015) In vivo...

...reliable and robust high quality viral vector service, making an array of ready-to-use viral vectors available... prioritization process to decide which tools we make available. This prioritization happens for good...of our most popular ready-to-use viral vectors, making up almost 10% of our requests by 2019 (Figure 5A...which preps to include in our service allows us to make ready-to-use viral preps that are available quickly...Hub before selecting an AAV can help a researcher make a more informed choice on which AAV to request and...since, we’ve learned so much about what it takes to make these viral vectors not only available, but accessible...

...to be a part of projects that connect people and make you work in a collaborative environment. Think of...conversations about your ideas and don’t be afraid to make something concrete happen. You may have more willing... institutions personally on conference calls and make sure to maintain these relationships by keeping ...organization. You can meet collaborators  for lunch, make a point of talking to them at a seminars, or even...from an internal to an external connection is to make sure you keep open lines of communication with colleagues...see interesting events, articles and newsletters I make sure to share them with a short note - this creates...

... Denotes the presence of an additional plasmid-- make sure to maintain this by growing on appropriate ...Induction timing is important, as you typically want to make sure your cells have first reached an appropriate...exponential growth phase are alive and healthy, which makes them ideal for protein expression. If you wait too... as there are not enough cells in the culture to make protein.  The DE3 lysogen/T7 promoter combination...positive selection and a p15A origin of replication, making it incompatible with other p15A plasmids. pLys ...Double-check your plasmid backbone and expression strain to make sure they are compatible. An arabinose-inducible...

... can promise is that my suggestions can help you make the best case for your candidacy. A major reason...complains that a seminar was too easy to understand. Make your seminar accessible. Let the experts ask you...minutes of the chalk talk. Realize that it is rare to make it through the details of all of the aims of a chalk...points you want to convey for each section or aim. Make sure those get priority. The fine details for aficionados...examples are not especially helpful. I set out to make something practical and accessible. If there's something...postdoc salaries all too well. If there's a group to make money from, postdocs are not that group. If you ...

...even one less run a week makes a big difference. Clean out freezers so you can make room and don’t have to..., check out my Recycling and Innovations posts. Making small changes in your daily habits can feel ineffective...sustainability problem, how can one little change make a difference? Individual changes to our behavior...wouldn’t leave your oven on all day to save time making dinner at night, so why leave your lab equipment...

...requires that the RNA be tagged. “This tool makes it possible to make the protein specific to any endogenous...PUF proteins! A newly available PUF Assembly Kit makes it possible to devise RNA binding proteins to hit...continue to evolve and improve, Abil says they will make those new plasmids available through Addgene, and...the final version,” she said. “If we learn how to make the library even better, we would be very happy ...

... puncta in each cell. When making figures for a paper or presentation, make sure the images are accessible...want to look for them? Or maybe you just want to make some really beautiful images? Multiplex IF may be...different colors in one session – fast and easy! Just make sure your primary antibodies are all of different...considerations to keep in mind. Most importantly, make sure you are using primary antibodies from different... and dye meets your needs and budget, or you can make your own. For example, to label the rat brain sample...

...people to take a break from their everyday tasks and make time for special projects? We borrowed an idea from...great success at accomplishing “we-should-really-make-time-for-that” type projects. Cross-team “Random...communication between labs. Scientists are often willing to make time for free food, and these lunches would provide...between labs, and people work hard at Addgene to make plasmid sharing as easy as possible. However, I ...work to share time with the people around us. It makes the quality of our work better and keeps us smiling...

... or set of plasmids. We hope that these articles make it easier for you to find and use the plasmids you...for in vivo expression, and/or produce vectors to make mutant cell lines or transgenic animals.   Wall...To expand the capabilities of Gibson assembly and make a more modular platform, the Jacks lab generated...cumbersome to use in a high throughput manner or make it difficult to quantify interaction strengths. ...Cas9-based AID tagging Seven years ago, Masato Kanemaki's lab successfully used the plant-based auxin-inducible...introduction of auxin range from 10-20 minutes, making AID an appealing tool for studying essential proteins...described in their recent Cell Reports paper, the Kanemaki lab overcame this hurdle using CRISPR technology...

... and 3' - G and CAGCT - 5' to the bottom oligo, making our final oligos 34 bp each: Top oligo: 5' - AATTC...

...appropriate water level in the water bath so before use, make sure that there is enough water inside....

...smallest bacteria. The absence of a rigid cell wall makes them resistant to antibiotics and antibacterial ...interpretation. Also, the signals from the nuclear area make the detection of mycoplasmas difficult (Ligasová...contamination when you are working without gloves or masks. Make sure to change the gloves and masks regularly instead...line stock for mycoplasma contamination before use. Make sure there is no contamination present in the serum...disposable pipettes to avoid cross-contamination. Make sure you keep all the required equipment and reagents...Avoid long exposure of the cell culture to air and make sure to tighten the flask cap before placing them...

...This is something I was expecting when I set out to make my first CRISPR/Cas9 gene edit. I wanted to inactivate...articles (gasp). To my delight, not only was I able to make the edits without any reagent-grade endangered Martian...Vinculin is a loading control. 3. In terms of making the monoclonal line: I had never done it before...important results (for example, if you needed to make a figure with the data). Using acrylamide instead...agarose to get sharper bands. Conclusion Overall, making a human cell line with a gene knockout was straightforward...who you are, you could quickly be on your way to making new scientific observations. References 1. Rago...

... an incredibly fast-moving field, and we want to make it easy for you to keep up with new developments...to update our web resources and plasmid pages to make them more useful and easy to navigate. We also updated.... Base Edit: Base editing plasmids allow you to make precise C:G->T:A or A:T->G:C edits in DNA without...Like RNA targeting systems, RNA editing does not make changes to the genome, thus providing an avenue ...CRISPR Genome Editing and The CRISPR Software Matchmaker Day 2: Homology directed repair: CRISPR 101: ...

... and then condensing that and making it accessible to students.” Making science approachable is often ...eScience Labs.  “We work with faculty members to make sure the courses encompass what they want to teach...onsite, Caitlin can visit the operations team to make sure “that everything that’s going into the kit ...non-profit Fred Hutchinson Cancer Research Center. And make sure to subscribe to the Career Channel of the blog...

...this responsibility seriously. Every decision we make, every project we undertake and every service we... that our materials are affordable but enough to make us self-sustaining. With this unique model, we are...opposed to fundraising.   Aside from enabling us to make all decisions with the goal of  benefiting our scientific...ever curious what specifically drives the decision making process at Addgene, just look to our core values...curated, high-quality data and reagents Addgene makes it easy for scientists to deposit and request plasmids...