...previously identified Cas13a-c orthologs, but they do include HEPN nuclease domains characteristic of the...here Find the Konermann et al. Plasmids here How do Cas13d enzymes compare to other Cas13s? Like other... and is thus HEPN-independent. These enzymes also do not require a protospacer flanking sequence, so you...

...Golden Gate etc.), and try to find the simplest way to do it (i.e. avoid assembling too many fragments at once...by methylation like XbaI, ClaI, etc. If you are doing multiple digestion be sure that the buffers and ...plasmid, and analyze it using the following steps: Do a single or double digest to be sure that the plasmid...

...groups and individuals with similar ideas to actually do something about it. Building a diverse team with ...There are many brilliant minds out there, but how do you get them to connect? Through networking of course...solving a community’s issue without involving them can do more harm than good. Check out this webinar for more...

...need to sort through them to find the desired clone. Do your selection, then screen Why add a screen to your...plasmids that contain the gRNA (white) from those that do not (blue). Let’s take a look at a few screening ... containing plasmids with inserts from those that do not is by using a restriction digest. The key is ...

...we understand what genes do! Viruses have several options to allow you to do exactly that: shRNAs are...clinical promise moving forward. What viruses can do for you Whether you are interested in basic science...long-lasting immune response even after a single dose. Vaccine development strategies. Image courtesy...

...inducible? How long do you need control? Will temporary control (a few days) be enough, or do you want a permanent...101: Repressible Promoters CRISPR 101: Which Cas9 Do I Choose for my Experiment Additional resources ...

...phenotype in the surviving population. Selections do require there to be a culture condition that can ...sound like a great shortcut, right? Who would ever do a screen when a selection can get you to the answer...culture condition which eliminates manipulations that do not affect the function of interest. An example where...culture cells in the drug of interest at a lethal dose for non-manipulated cells and then assay cells which...

...” of mentors as one key to their success. But how do you find these elusive teachers, supporters and advisors...be read or watched during the meeting.  Scientists do better with process, so don’t just get together and...blog in this series will have more ideas on what to do once you are in a mentoring relationship.  Celebrate...

...opportunities to fall in your lap. Sometimes, in order to do the thing you love, you have to create opportunities...honest), I would tell them to follow your passion. Do what you love. Don't worry about the rest.”- Jason...

...maybe smaller Cas systems could exist in nature. To do this, they mined a database of microbial genome and...indiscriminately cleave ssDNA in trans like Cas13 and Cas12 do for RNA and dsDNA, respectively. However the authors...

...Unfortunately, many science graduate programs are still doing an inadequate job of supporting science trainee ...into diverse, successful careers.  We’re not even doing a very good job of providing skills training for...

...Addgene, so we need to be cost and time efficient. We do this by making our own competent cells and using ...each new batch of competent cells, don’t forget to do a negative control (plate your cells without adding...

... plans to raise salaries. What are institutions doing after the injunction? We are particularly interested... have found out about what their institution is doing from Future of Research over social media, not from...recommendations, even when a federal mandate encourages them to do so. Immediately after the injunction, as we started...

...procedure dramatically reduces the number of clones that do not contain the recombinant plasmid and therefore...are identified much more efficiently, as those that do not contain the desired insert should not grow.   ...be present. Most common cloning strains of E. coli do not contain the F plasmid (and are considered F-)...

...cells - a rich soup of life encircling the tumor. How do microenvironment enzymes and metabolic reprograming...mediators and improving cancer cells proliferation.  How do microenvironments affect tumor spread and drug resistance...for a tumor to spread and to resist drugs. And to do that, let’s take a look at exosomes. Proficient in...

...without a kit that outlined several reasons why doing something without a kit has advantages: less plastic...without saying that one should take care whenever doing any type of DNA or RNA purification to avoid contamination...expression. There are several things that researchers can do to combat this issue. First and foremost, be mindful...

...you have all this knowledge, the question is: How do you know what’s too much (or too little)?  The answer...things aren't working out, adjust the ask.  I used to do a lot of outreach to student groups and being aware...think about everything you’re asking your learners to do, it becomes a lot easier to build successful, accessible...

...fill this gap by both highlighting that researchers do learn MANY transferable skills while working in the...just a question you have about the world. You might do some background research to better understand it ...method to problems in a variety of contexts.  How do you develop problem solving skills? Thankfully, as...

...of proteins functioning in a variety of ways. How do scientists focus on the activity or function of a...plasmids here! Where should I place the tag and how many do I need? When designing a plasmid containing a polyhistidine...perform assays that they otherwise would not be able to do. We hope that this blog has answered all of your ...

... synthetic modifications, specifically ones which do not contain a 5’ triphosphate in the gRNA, reduce...many suitable dyes to choose from and you can opt to do-it-yourself with a kit or purchase it commercially...but you can still choose what dye to use  What to do once you’ve chosen a gRNA and modification(s) Congratulations...