...single molecules and newly formed proteins.  Limitations with GFP in imaging  While GFP has been an immensely...to study proteins in vivo, it does have some limitations. GFP has long protein maturation times which ...antibody based probes a solution? To address these limitations, scientists have co-opted antibodies for tagging... the National Academy of Sciences of the United States of America, 69(9), 2659–2662. PubMed PMID:4560694...

...choice while bypassing the intermediate pluripotent state. Though a valuable tool for regenerative medicine...least day 13. However, distinct transcriptional states were seen as early as day 6. All together these...stochastic, but rather there’s a particular cell state that’s permissive to reprogramming.In this lineage...reprogramming. MettI7a1 encodes an uncharacterized putative methyltransferase. Adding Mettl7a1 to the standard...

...however, has inherent limitations because it turns a dynamic process into a static measurement and it provides... acidic environments at 586 nm. Regardless of excitation wavelength, Keima emits at 620 nm. The ratio ... Noda, Takeshi, and Daniel J. Klionsky. "The quantitative Pho8Δ60 assay of nonspecific autophagy." Methods...

... sketch on the board without leaving Zoom. The annotate function can also make the screen-sharing experience...top of the screen and every participant get to annotate things as they come up in their minds. The final...writing and this recent article published in PLOS Computational Biology (Frassl et al., 2018): “Science is increasingly...where all lab members talk no science. No data presentation will appear here, and only general chatter will...

...train and acquire the data you need on the new instrumentation. Bonus reasons Visiting gives you access to...This probably isn’t the lab’s first request for visitation if they have specialized skills and equipment...so that you can work together to ensure your expectations are reasonable, your reagents are onsite before...include, but is not limited to temporary housing, transportation, living expenses, and necessary reagents. Your...

...forming concatemers later. The "FS" stands for Fragmentation System and this is the most important part of...for library preparation using this kit is: Fragmentation and end repair Adapter Ligation Clean up/Size... the start of the workshop, NEB gave a brief presentation describing the technology. Then we split into...and network with other technicians/company representatives. I got to talk with a technician from Neon ...

... gRNA representation, produce high titer lentivirus from the library, ensure gRNA representation in the...technologies in plasmid form and are proud to have facilitated their fast adoption. However, in many cases the...screening-scale experiment achieving the gRNA representation recommended in the publications associated ...

...success. It is imperative that you resist the temptation to “just do it myself” or micromanage –you don...develop the web page. The team manager must then facilitate appropriate communication between members of ...only way and may not even be the best way. This facilitates success and trust. Micromanaging is bad for everyone..., meeting minutes and plan changes. Written documentation helps make sure everyone is on the same page...

...samples, the rest will be put into a status we call “Archived.” This status allows us to store plasmids and..., please fill out this form and we’ll add your citation to the plasmid page on our website. Contributing... contain the expected features and are free of mutation.  Once the samples arrive, each group of plates...

...effective manager. Mismanaging (no pun intended) expectations —yours and your team's—can lead to frustration...unnatural for those accustomed to hands-on work. The temptation to "do it yourself" can result in burnout and...Miscommunication often arises when managers fail to set expectations or provide timely feedback, leading to unproductive...discussions to understand their perspectives and facilitated a team meeting to highlight their complementary...

... used FRET, but it eliminates the need for an excitation light source. Instead, a luciferase substrate...complement optogenetic tools, where light used for excitation can cause unintended activation of the optogenetic...various colors. In fluorescent imaging, you use the excitation wavelength to specify which FP emission you’ll...Nano-lantern system does not use external light for excitation, you instead have to rely on differences in emission...

... methods of manipulation are genetic knockout mutations and treatment with small molecules/drugs. With...with a uniform population possessing a specific mutation or phenotype. For example, a drug screen may be... performed on a cell line possessing a patient mutation to assess how patients may respond to drug treatment...survive as these must have acquired a favorable mutation. But if the cellular pathway of interest conveys...

...collaborating as they craft sentences for abstracts and presentation slides, learn from audiences as they do science...something they did not understand about your presentation, question whether other aspects of the work ...primary focus When writing a paper or preparing a presentation, it's easy to forget that you have a much bigger...practice.  Ask them what they learned from your presentation and evaluate whether or not you’ve succeeded...

...for NHEJ repair regardless of end structure, and dictate the major events of the pathway: Broken ends are...processing of DNA ends tends to be the point where mutations are introduced. Repair of Cas9-induced breaks ...about a two-thirds chance of causing a frameshift mutation. Of some importance, the deletion can be less ...double nickase” strategy vastly reduces breaks and mutations at off-target sites. Christina Mork contributed...

...variation of gene expression. Because pSC101 is not representative of all types of plasmids, the stabilized promoters... number. During the shift from exponential to stationary phase, the number of plasmids per cell increases... plasmid copy number during the transition to stationary phase, Segall-Shapiro et al. placed a trans-acting... such as qPCR or microarray. But, aside from facilitating the joys of a p-value < 0.05, having such fine...

...times, LinkedIn may revoke your ability to send invitations. Finally, if you don’t know clearly why you are... box, it will send an unpersonalized, generic invitation. Don’t try to connect from the LinkedIn phone... app either as this also results in a generic invitation. 3) Connect with people you have really talked... with this and as long as you personalize the invitation with your intention for connecting, this is probably...

...Figure 1: Overview of using Sandwich ELISA to quantitatively measure protein concentration or antibody specificity...Figure 2: Overview of using Competitive ELISA to quantitatively measure protein concentration or antibody specificity...Figure 3: Overview of using Direct ELISA to quantitatively measure protein concentration or antibody specificity...Figure 4: Overview of using Indirect ELISA to quantitatively measure protein concentration or antibody specificity...

...understanding of how our bodies work, new treatments for debilitating diseases, and maybe even ways to alleviate health... broaden the conversations about genetics and facilitate dialogue between scientists and communities. ...With our focus on engaging the community and facilitating dialogue, a major part of our everyday work ...jargon-filled language in academic writing and presentations. But for many people who don’t work in these...

...consequences of that change. For example, “Mutated aspartate 123 to Lysine (D123K), lowers biosensor affinity...indicated in the “Species Other” column. Relevant Mutations Indicate the amino acid change as well as any...accept sequence files in any format. Distribution status for your plasmids: Hold for Publication or Distribute...

...turn. The first is the passing and upcoming implementation of the Federal Labor Standards Act (FLSA) which... in salary, particularly given that this has precipitated increases to the NIH recommended pay scale –...and opportunities for us all.  For example, implementation and compliance for the FLSA changes, including...challenging, but will be aided by the development and implementation of new strategies designed to cope with current...

...after amputation. For example, the dramatic “two-headed” phenotype is observable in post-amputation planarians... experiments, as well as how to perform worm amputations. They also created an online companion resource...beta-catenin. Other gene targets do not require amputation to be visible. For example, odf2 gene knockdown...

... A major drawback of this method is the stray excitation of neighboring neurons in opsin dense regions...therapeutic methylxanthine drugs like theophylline. To facilitate future work with this goal, the Barrick lab has...N-demethylases and other required enzymes, including a glutathione S-transferase homolog from a Janthinobacterium...dark yellow fluorescent protein (YFP) with an excitation peak of 519 nm. These proteins have improved ...

... NPA states that Postdoctoral scholars should have the skills and techniques needed to facilitate effective... Chair automatically as it was an understood expectation of any Vice-Chair. Following an official assessment...leader in the NPA, I had to speak as an NPA representative and speak the NPA’s stance. I had to be professional...

...approach for harvesting my AAV. I skip the PEG precipitation of the HEK293 cell culture supernatant because...pellet. My yield would be higher if I used PEG precipitation, but for my experiments, I get enough virus ...culture media, you should consider performing PEG precipitation. Instead of sonicating the cell pellet, I lyse...with this AAV lysis buffer and freeze/thaw the lystate four times in a 95% ethanol and dry ice bath to...

... However, safety requirements and expression limitations may be too constraining for some experiments,...in your experiment. Figure 2: Visual representation of common AAV serotypes used for different ...that reside outside a host cell’s genome. In this state, AAV cargo can be stably expressed in non-dividing... on the Addgene Blog Overcoming the AAV Size Limitation for CRISPR Delivery Genome-wide Screening Using...

...accounting for some of the inherent inaccuracies of computational prediction models. In addition, we provide options...most promising pegRNA designs and especially facilitates the use of prime editing in pooled settings with...models (such as PRIDICT/PRIDICT2.0) can greatly facilitate prime editing experiments, thus saving you time...Switzerland), where he developed a strong passion for computational biology. His expertise includes applying machine...

...molecular biology and bioengineering, as it has facilitated the development of a simple and scalable means... gene expression. Its catalytic domains can be mutated to inactivate the protein’s nucleolytic capability...interrogation of gene function. However, there are limitations to a system that relies on two distinct Cas9 ...possible, this strategy suffers from several limitations. Perhaps most importantly, most Cas9 orthologs...

... or C terminal of a protein and can inducibly facilitate the degradation of the tagged protein. Degrons...here are all conditional, meaning that they only facilitate degradation of their fused protein under certain...through interacting ligands. After an E3 ligase facilitates ubiquitination of the Halo-tagged protein in ...HaloTag and its PROTAC are all that is needed to facilitate degradation. HaloTag is also the degron tag with...

...using the MoClo system. The paper also gives quantitative data for regulatory elements, which has not ...example, knock out a gene with simultaneous complementation, with a knockout and expression construct on...tools, double dimerization GFPs, and circular permutation GFPs. Hopefully we will have new tools for plant...doing. They look at structure, impediments, the representation of women on committees, and how the culture...

...like guanine. This RNA editor would permit point mutations in RNA which could recapitulate or rescue known...REPAIRv1, the lab targeted 34 pathogenic G->A mutations from the ClinVar database. They successfully edited...still displays substantial off-target activity. By mutating residues in ADAR2 that interact with duplex RNA...rational mutagenesis and identified that a S375A mutation in RESCUE (named RESCUE-S) offered ~76% on-target...