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...experiments? While codon preferences can be very useful for organisms, they can be problematic for researchers...organisms - organisms have codon usage bias. You can find many codon tables showing which codons encode...constraints that lead to these preferences and what can we do about them? Read on to find out!   Why do organisms...conditions. With varied codon usage, an organism can change which proteins are highly expressed and which...instance, it may not express at all in E. coli (you can find a variety of databases showing various organisms...sounds. Even for relatively short peptides, there can be many possible ways to encode them and what constitutes...advantage of producing additional tRNAs is that you can use the same expression system for many different...

...The analysis can be sorted by any of the parameters displayed in the summary table. You can also use your...analysis formats: sample by sample analysis, which can compare up to five editing experiments at a time,...simultaneously.  The current version of ICE software can analyze indels that result from single or multiplex...red exclamation point in front of that sample. You can hover over the yellow or red circles to gather details...  The higher the R2 value, the more confident you can be in the ICE score. KO Score - Represents the proportion...to search for a particular sequence or name. You can perform more in-depth analyses on each sample by ...about the indel profile of the edited sample. You can also download the entire analysis as a ZIP file by...

...although sequences with G/C content outside this range can still be optimized (Anzalone et al., 2019). Test ... pegRNAs whose 3′ extension begins with a C base can cause unwanted base pairing (left), while pegRNAs... What to edit The sequence you choose to install can influence the success of your prime edit. If possible...synthesized strand before heteroduplex resolution, which can lead to indels (Anzalone et al., 2019). Figure...left), but “bubbles” of 3 or more mismatched bases can often evade MMR (right). Created with BioRender.com...PE3/PE5 approach. Figure 7: Nicking sgRNAs can be designed to target edited DNA, so that nicking...scaffold sequence (orange) and the genomic target can increase the odds of incorrect editing outcomes, ...

...overhangs, stat? GetSet can do that for you, complete with predicted fidelity. You can specify number of overhangs...highest-fidelity overhang set it can find for those parameters. For even more specificity, you can exclude specific sequences...overhangs with a high potential for mismatches, so you can redesign individual pieces as needed to improve fidelity...rather than pre-designed sets, so identical searches can return multiple answers. You might want to take advantage...fusion sites to split it into multiple fragments. You can define the number of fragments, and the “search windows...constricted to a specific number of fragments, you can also ask it to give the maximum or minimum number...52 fragments may be near the upper limit of what can be accomplished with Golden Gate Assembly—even when...

...region or nucleus. Just optimizing AAV expression can take months (or longer), yet the process is often...detailed in the primary literature, and as a result can leave scientists under-informed with respect to how...cell types used for packaging viral particles (6), can influence vector performance. Open science initiatives...especially for AAV data, that may not be the case. We can all acknowledge that negative data is something every...getting this information out into the public where it can be utilized by others. Addgene’s AAV Data Hub is ...evaluate how small changes in vector administration can influence AAV expression properties.  By providing...generally captured in the literature, scientists can use the Data Hub to compare different methods and...

... that you can do now is diversify. Read on for an overview of some of the resources you can use to better...Conferences One of the single most important things you can do to plan for your career is to expose yourself ...homework and know who is going to be there, so that you can tailor your CV accordingly. Make it stand out with...you’ve identified an appropriate job opening, you can send your contact an email with the relevant info... position he or she originally called about, you can keep in touch and reach back out when you are looking...should have a well-groomed LinkedIn page. Here you can search for jobs, post your CV, and, most importantly...who they know all in one place! This information can be critical to a focused and effective job search...

...visualization for big biology. GPS for the genome We can think of the human genome as a map with three coordinates...addresses in the genome. Let’s now examine how we can locate powerful stretches of DNA known as enhancers...Through the marvel of DNA compaction, an enhancer can increase the expression of a gene that is millions...are bound to modified proteins called histones. We can detect modified histones with “GPS for proteins,”...tri-methylated histones, functions as an enhancer. We can therefore denote enhancers as follows: (mono_methylation... and conclude with a targeted CRISPR screen that can disrupt these enhancers. A real-world experiment ...H3K4me3 The following video demonstrates how anyone can find enhancers with Quilt. Seek and destroy enhancers...

...Gal4/UAS system, and you can read more about them in review articles (2–4). You can also take a look at my...expressed when Gal4 is expressed. Gal4 expression can be regulated by any chosen promoter, and Bloomington...expressing Gal4 in your tissue type of interest. You can quickly mate your UAS-YFG line with a variety of ... next blog post). For example, the hedgehog gene can be overexpressed in several cell types by using different...Controlling multiple genes with Gal4/UAS system One can also use Gal4 to drive expression of multiple genes...’s not always clear which side he roots for. You can try to convince him to use Twitter @jonchowphd but...

...metabolism of leaves can be changed by light and temperature and leaf autofluorescence can interfere with microscopy...that replicate current knowledge, but the sensor can also kinetically outperform current methods. Unger...nucleotide polymorphisms present in the human population can affect drug and dose response. Selective serotonin...Unger ultimately hopes that medical professionals can genotype the patient and rapidly test the variants...promoters responsive to water or nutrient levels can help him determine whether the artificial leaf behaves...labelled with a constitutively fluorescent reporter can also be used to track how one population responds...Mitja Remus-Emsermann’s lab at the University of Canterbury where he studies how bacteria interact with each...

...yours and your team's—can lead to frustration. Clinging to technical tasksDelegation can feel unnatural for...contemplating the leap into management, the journey can be as exciting as it is challenging. How do you... you need to know.   Figure 1: Management can seem a mystery at first! Image used under CC.  ...hands-on work. The temptation to "do it yourself" can result in burnout and underutilized team potential...connecting with mentors who have made similar transitions can provide you with insights, lessons from their experiences... Myers-Briggs Type Indicator or DISC assessments can help you navigate team dynamics effectively and foster... The Making of a Manager by Julie Zhuo Radical Candor by Kim Scott Courses: Leadership programs on...

...grants and projects that might help your work Tenure can depend on your national recognition and the more ...academia is all driven by personal connections What can you do to get the most from the time and money you...conference. Large conferences have many opportunities but can be overwhelming as far as meeting new people. At ... want to introduce yourself and what you do. You can probably use some scientific terms when socializing... that already). Start with, “Is this seat taken? Can I join you?” Next talk about the food and then move...up to someone much more senior than you, but you can safely seek out scientists your own age. Some will...LinkedIn users notice this sort of connection. You can also see and offer congratulations on job changes...

... determines which tissues and cells an AAV can infect) can influence an AAV’s value greatly. Applying ...have different tropisms but, as you can see, AAVs of each serotype can transduce multiple tissues. So how...transducing different brain regions. Although AAV2 can transduce many tissue types, it has low expression... It has been demonstrated that certain serotypes can undergo axoplasmic transport. These AAVs may move... If axonal transport is not desirable, transport can be reduced by injecting a smaller dose of AAV and... than other viruses, multiple freeze thaw cycles can still reduce AAV titer. For long term storage, make... begins. ScAAV avoid this rate limiting step and can immediately undergo replication and transcription...

...Figure 1). Antibodies can be broken up into smaller parts, the variable regions can be altered for specific...If you think functionality can be ramped up even more, the best mutant can then be used as the starting...delivering other treatments, their efficacy can be significantly increased through structural and functional...specific functions, the constant regions can be altered for different isotypes, and so much more. How you...low-fidelity polymerase, like Taq. Other PCR conditions can be changed to further lower fidelity, like changing... and the dNTP ratio. The resulting random errors can give you new and unexpected sequences. Degenerate...mutate whole antibody sequences, degenerate primers can be designed to selectively mutate specific regions...

...Career and skills development can happen anywhere, whether you realize it or not. In the lab, you could...two projects: one “safe” project that you know you can get a publication with and one “shoot for the moon...you don’t think you’re qualified. The worst that can happen is they say no, which is infinitely better...Senior Software Engineer “Searching for a new job can be an imposing endeavor.  Break the process down ...Get the toughest steps out of the way early if you can.” - Eddy Page, Digital Media Production Specialist...Kamens, Executive Director Career exploration “You can't wait for opportunities to fall in your lap. Sometimes...

...which level of purity. Based on these premises we can classify a plasmid preparation in 3 different ways...electrocompetent cells is minimal (picogram range). It can be extracted from small bacterial cultures (i.e. ...prokaryotes or eukaryotes into a synthetic vector that can have a plasmid or viral origin. The type of cloning... extraction. Restriction enzyme based approaches can require several hundreds of nanograms or micrograms...you want to utilize your plasmid DNA, so that you can decide what will be the optimal type of plasmid preparation...

... – Get your voice out there Here’s one thing you can do today: Sign up for a Twitter account. Check out...in local museums, zoos or botanical gardens. You can also work or volunteer for a science communication...you don’t have time to maintain your own blog, you can probably write for your institute’s blog. The Wellcome...Use your unique talent to communicate science. You can draw live sketches at conferences like @ATJCagan ... by taking a media or journalism course. Courses can help you gain work experience and build your network... expect them to be the “be-all-and-end-all”. You can, for example, get an MSc Science Communication or...involve the public in your research projects, you can set up a citizen science project. Answering the big...

...constructs can be achieved by fluorescence-activated cell sorting, and stable gRNA expression can be achieved... But other enzymatic fusions, such as dCas9-TET, can, like the iron ball-and-chain that they resemble,...site is not as straightforward as expected. TET1 can have many effects independent of dCas9 targeting,...ve got some good news: complete DNA demethylation can be achieved by dCas9 alone, without TET (or any demethylase...demethylase) fusion. This may seem counterintuitive: how can a DNA-binding protein with no enzymatic activity ...DNA demethylation! With optimization, this method can produce nearly 100% demethylation without the confounds...block that unmethylated CpG so that nothing else can interact with it. Remember, the goal of this experiment...

...hybridoma cell lines can produce more than one mAb (Bradbury et al., 2018). Hybridomas can lose gene expression...into a fully functional antibody. These antibodies can be used just as you would use antibodies made from...purified antibodies from a patient’s blood), scientists can use mass spectrometry to determine the amino acid...from an established hybridoma line, the antibody can be converted into recombinant form through DNA sequencing...2010; Andrews et al., 2019). Finally, scientists can perform the entire process of antibody selection ...been cloned into an expression plasmid, the plasmid can be introduced into host cells, such as bacterial,...engineer them. Given the ease with which scientists can now improve genetically-encoded research tools, it...

...what facet of academia you are in, grant writing can be the lynch pin that results in your success or ..., and perhaps the most important bit of advice I can suggest, is to give yourself plenty of time when ...and why these findings will be worth knowing? This can sometimes be tricky, as most of our research builds...collaborate with specialists in different fields that can provide different insight into an area - not just...Furthermore, using new technologies to solve a problem can allow you to approach that problem in a novel way...results published in a paper, but positive outcomes can include poster presentations, talks about your work...grant application to your peers and collaborators can also help you get some critical insight into what...

... tools can distinguish between global strains of the Zika virus (Asian-, African- and American-lineages...monitor and slow this spread until vaccine programs can be put in place to provide community protection. ...allow for the rational design of RNA sensors that can detect virtually any RNA sequence (Green et al, 2014... later. The Zika publication timeline I think we can speak for all the authors on the paper when we say... of a rapid design and prototyping pipeline that can take pathogen sequence information from public databases...technology on active virus, as well as shown that we can detect the virus in a clinically relevant setting...patient blood and urine samples; we showed that we can detect the virus directly from infected monkey blood...