We narrowed to 821 results for: EED

...target cells and the desired edits. First, you will need to know what the ploidy of your target site is. ...line, so it’s important to verify that you won’t need to screen hundreds of clones looking for your edit...This way you can ensure you aren’t looking for a needle in a haystack before going on to generate and screen...then a single, bulk validation may be enough to proceed with your experimental plan. Validating knockout...generate your knockout? Then you’re in luck! You don’t need to sequence your PCR to visualize an edit. Most ...clones, however, sequence confirmation is still needed to verify the correct sequence was inserted or ... edited cells. As with all other methods, you’ll need to further validate the specific mutations using...

... and by offering advice on areas where you might need some help. Today in our transferable skills guide...something like, “I’ll be there in a few minutes, I just need to split some cells.” Inevitably, this task would...techniques of your own, but if you ever find yourself in need of a few ideas, I hope you’ll find this post useful...the energy to finish any additional work you might need to do. In most cases, I would recommend the second...outset of a task, I outline the activities that are needed to accomplish these goals. Practically speaking... remind myself of the specific activities that I need to work on. Throughout the day, I cross items off...they are accomplished or I add to the list as the need arises.Why do I write things down? It’s more viscerally...

...pieces of a 2,000 piece puzzle. A scientist would need to run potentially hundreds of Sanger sequencing...Sequence FLAG DYKDDDDK HA YPYDVPDYA His HHHHHH Myc EQKLISEEDL V5 GKPIPNPLLGLDST Xpress DLDDDDK or DLYDDDDK ...

... into the PX330 vector backbone? A2: There is no need to add the NGG PAM sequence. Because the 'NGG' of...the PAM is used to select your genomic target, you need to make sure the NGG immediately follows your target...should order is:5'-CACCGN...N-3'3'-CN...NCAAA-5'You need to add the CACC and AAAC cloning overhang. Q3: When...starts with a 'G', you naturally have one and do not need to add an additional 'G'. Q4: Where are double strand...be sufficient to generate off-target DSBs; there needs to be extensive homology of the protospacer followed...your cells. If you want to use double nickase, you need to have four spacers co-expressed and using PX335...the double nickase system, with PX335 vector, you need to find two guides for each cleavage site, but this...

...Fluorescent proteins are very stable in SeeDB2. However, SeeDB2 is not powerful enough for large brain ...2014), before clearing with SeeDB2. For more information, visit our SeeDB Resources website. We update...proteins were best imaged after tissue clearing with SeeDB2 (Ke et al., 2016, 2018).   How to use Tetbow for... for High-resolution fluorescence imaging using SeeDB2." Bio-Protocol 8 (2018): e3046. Livet, Jean, et...Ultrafast tissue staining with chemical tags." Proceedings of the National Academy of Sciences111.36 (2014...

...classmates! Your mentors don’t need to be more senior or more “important”, they just need to know you well enough...you are more desperate or needy so you can’t wait until you imminently need a job. Start now. Informational... group We don’t all get the mentoring support we need, and most of us could use more mentors to listen...any science career, there will be a time when you need to speak publicly to communicate your work. One ...career options. 5. Write a grant - even if you don’t need one A lot of science in academia and not in academia...

...may only need to validate on denatured proteins or on live cells. But for others, you may need to validate...well-organized, documented, and easy to find (though we might need to see it to believe it.) As you browse the collection... affect its configuration. You don't necessarily need to know where the epitopes are, just if the available...selected an antibody, or multiple antibodies, you’ll need to validate them for your protein and its configuration...configuration in your assay. For some assays, you may only need to validate for recognizing the protein following...designing your controls in validation, as you may need to test different configurations and conditions ...

...individual target species. Indeed, in order to use MAGE, the λ Red recombinase enzymes need to be expressed and... easier for you to find and use the plasmids you need. You can find all the hot plasmids from 2016 below... kit Synthetic and basic biologists alike often need to test multiple expression systems before they ...digest and Sanger sequencing, respectively. The speed and simplicity of GMAP allows scientists to easily...native methyl-directed mismatch repair (MMR) system needs to be repressed in the host strain. To address this...variety of new compounds being generated, there is a need for genetic optimization of industrial yeast strains...supplied as annealed synthetic oligos - eliminating the need for PCR or additional molecular cloning steps during...

...polymerize, allowing the slower exonuclease activity to proceed. By including only one type of dNTP in the reaction... can decide exactly how far the T4 exonuclease proceeds: at the first occurrence of the chosen nucleotide...if those overhangs perfectly match up. They just need enough complementary ssDNA to hold the plasmid together...ways. One option, imprecise T4 digestion (no dNTPs needed), allows both insert and vector to be digested ...incomplete PCR — although higher DNA concentrations are needed for this method to be successful. Low DNA concentration...approaches require higher concentrations of insert to succeed. 5′ overhangs are generated in the linearized vector... you can assemble multiple fragments without the need for specific restriction sites or DNA ligase! This...

...external stakeholders to learn more about their needs and any problems that they are trying to solve. ...test the solutions with the users, gather more feedback, and start the cycle over again to refine the ...software possible. Similarly, when the developers needed to know information from the lab folks, it was ...with our software or devices, and anything they may need to help improve their work. Then, I distill and ... well as the developers. Lastly, you’re going to need to be a problem-solver. A lot of the time, the problem...clear, but the solution is more complicated. You’ll need to be creative to be able to work with others to...strong relationship-building and influence. You’ll need a large deal of influence and strong relationship-building...

...answer the question, you need only to take that step. Repeat that as often as needed until you get to the ...calm: “Excuse me, I seem to have lost my place. I need a minute.” I then pulled out my notebook, opened...presentation will be better if you take time when you need it, rather than trying to push through a state of... deck gets uploaded. Your mic emits screeching feedback. You’re muted on Zoom. Your video conferencing...telling yourself, “I can stop and check my notes if I need to,” or “It is okay if it doesn’t go perfectly,”...group of peers who are there to provide critical feedback, often with a semi-international audience or not...

...it off? Turn it down, but not forever? You don’t need to resort to the force to control protein expression...off, tunable, and/or inducible? How long do you need control? Will temporary control (a few days) be ...interest. CRISPR-Cas9 and a custom gRNA is all you need to generate a frameshift mutation that will permanently...permanently ablate protein expression. If you need the ability to control when you turn off expression ...you would like. Check out our promoter blog if you need help choosing. Viral expression is another mechanism...exogenously introduced proteins, as many of them just need a special promoter or tag prior to introduction ...endogenously expressed protein, then these systems need to be site-specifically knocked in. This can be ...

...yeast cells. Need more MoClo? Check out Addgene's MoClo Guide! Customizing MoClo for your needs As you probably...read and then come back here once you are up to speed on the basics of the technology. In this blog, we...benefit from a larger library of parts but don’t need to add tags to your proteins, then the CIDAR Kit...a MoClo kit plus the plant-specific parts you‘ll need, including sub-cellular localization signals, plant...It is important to understand your experimental needs before choosing one or more of these kits, as there...specifically for the types of expression and integration needed in mammalian cell types, and comes with a large... to select an available kit that best fits your needs and then modify it as necessary. And remember, if...

... style according to the needs of my committee members. Some members would need much more guidance, direction... scholars should have the skills and techniques needed to facilitate effective teamwork, manage day to...sticking to deadlines and performing the work I agreed upon (just as I would in my laboratory work). Committee...would act more independently and some others would need regular re-direction and alignment with the overall...responsibilities Being a leader in an organization, you need to make sure the organization speaks in “one voice... you can find opportunities around you. You just need to be prepared to take on the challenges and be ...

...then need to be purified by density gradient centrifugation (e.g., iodixanol) using a high-speed ultracentrifuge...cells for further processing. What component(s) you need to collect will depend on the serotype, as some ...however, you’ve collected the producer cells, you’ll need to lyse the cells to let the virions out. We recommend...exchange and further concentrate the virions if needed. For more details on the production process, check...produced, harvested, and purified your rAAV, you’ll need to determine the titer of your viral vector solution...).  To choose the optimal titering assay, you’ll need to understand how different titers are measured....

...increase plasmid yield. Keep in mind that you might need to use multiple techniques for a particularly stubborn...bacterial strain to see if additional steps are needed to optimize yield. You might want to include an... high levels of protein, not plasmid DNA. If you need large amounts of DNA from a plasmid in a protein...optimize your growth time, temperature, and shaking speed to maximize cell density and plasmid yield as these...hours, but cultures with lower copy plasmids may need to be grown for 20 hours or more to achieve maximum...culture volume and you should ensure the shaker speed is fast enough for sufficient gas exchange in the...those in deep-well plates, require a faster shaking speed for proper aeration (260-300 RPM). 37℃ doesn’t work...

...What is a transferable skill? Time management. I needed it when balancing a handful of demanding courses...work, bills, (at times) a social life and rest. I need it just as much in my current role as Customer Support...to do the job. You may think this means you’re guaranteed to get the job, however, many times the hiring...your boss approaches your team with a project she needs one of you to lead. At first you might think, “I...change happens at your organization, when your boss needs someone to step up, when a new venture presents ... Say your manager approaches you with glowing feedback from a client you helped. Maybe you awkwardly ...other things you have to do. Pause! Enjoy that feedback, bask in the glow of a job well done, take some...

...get this far and realize you need to apply different filters you do not need to start a new search. Simply...can narrow your search results to exactly what you need: Better searching with filters Not every plasmid...filters lets you drill down to the exact plasmid you need. The ability to apply a wide range of filters isn... your search results to match your experimental needs. Remember, only filters with results will show up...Serotype: Choose the AAV capsid or lentiviral prep needed Tags: Identify a specific marker protein Use: Similar...website and we hope it leads you to exactly what you need with every use. If it doesn’t and you find yourself...

...review. If it’s a well-researched field, you may need to get specific to make sure your article doesn’...reviews to get a list of research papers that I needed to read. Once I had an idea of the themes in the...word of advice: when reading, don’t feel like you need to read every paper from start to finish. Find the... comment about writing: initially I felt that I needed a huge chunk of time to sit down and get anything...telling the story. Like any editing process, you will need time away from the article to be able to keep editing...normal and it means you’re on the right track!  Get feedback After you’re done editing, send it to some actual...actual experts in the field for their feedback on the scope and the content. Submit your article! Overall...

... thank Seeding Labs for this continued opportunity to get plasmids to the researchers who need them the...Sciences in Ghana will be receiving the third Addgene/Seeding Labs plasmid grant and plans to use the grant to...Future awardees We will be opening the Addgene/Seeding Labs Plasmid Grant to a third round of applicants... seeding labs... seeding labs plasmid grant...