...go about starting a lab? Obviously, funding and support are crucial parameters, and familiarity with equipment...a PhD student in his first year. “It was very important to me to be with them in the lab when they started...spending more time in his office, he gradually incorporated regular group meetings, journal clubs, and focused...the lab and to take things slowly! It is more important to carefully select first hires who are well-suited...seek out advice from seasoned PIs who can offer support. “It is an exciting and emotional time!” George...a challenging process. Each PI emphasizes the importance of slowly building up your lab and selectively...

...strategies for finding what they need, and we wanted to support all of them — searching, browsing, drilling down...experts, our software team tailored Elasticsearch to support the different usage patterns the larger team identified... selecting a suggestion — how should these be supported and combined?  Luckily, we had the advantage of... information in our search indices in order to support different kinds of matches: exact words, partial... terms.  Weighting We determined the relative importance of different bits of information for a given ...that make our new search engine unique. Most importantly, with this new implementation, we laid the foundations...

...off for the day. 1:45 PM--Write final report. Work on a final report for a user research project I led. ...field-based software product manager at Oxford Nanopore Technologies. If the first thing you hear when...streamlined manner. Now, in my current role at Oxford Nanopore Technologies, I work with our customers in the...believe that more and more people will have the opportunity to explore a career in product management.  A...currently a Field-based Product Manager at Oxford Nanopore Technologies for the North America region. She...

...individual cells. Electroporation on the other hand uses electrical pulses to create transient pores in the cell...plasmid of interest. Scientists can also use electroporation, the application of an electrical charge to...lysogenic cycle, the phage’s genetic material is incorporated into the host’s genome at a particular integration... and stable cell lines, where foreign DNA is incorporated into the cell’s genome and is thus passed down...charged cell membrane. Physical methods such as electroporation or microinjection actually pokes holes in the...

...increased complexity comes the need (and/or opportunities) to modify the device for individual experiments...openUC2, which I am a co-founder of), that aim to support the community in developing better, more sustainable...difficult to overcome. Universities may not have the opportunity to buy parts through low-cost sources such as...accessibility. Through this approach, the Planktoscope, a portable device for studying plankton using flow cells,...quickly with the printer, act here as a kind of "teleporter," since a part developed in Germany, for example...-based models and the use of taxpayer money to support this research. But there is a move towards more...

...phosphatase-positive assay) as well as stabilization (Nanog-GFP reporter activation and silencing of factor-linked mCherry...are available at Addgene. Kim, et al. Stem Cell Reports. 2015. PubMed PMID: 25772473 Targeting and manipulating...Genetically modified organisms like transgenic mice are important tools for understanding the function of cell types...enhancer elements and combined it with a variety of reporters (i.e. eGFP) and effectors (i.e. DREADDs). The ...CRISPR edits and improves editing efficiency. Importantly, the point mutations created by Agudelo et al...et al. also mutagenized the cytidine deaminase portion of the base editor to create SpCas9 base editors...reagents shared via Addgene. Narayan S, et al. Cell Reports. 2017. PubMed PMID: 28813671 Listen to the podcast...

...Harvard University. No matter whether you are a sports photographer at the Super Bowl, a medical technologist...the first fluorescence microscopy technique to incorporate optical sectioning. Optical sectioning refers...their fast movements. Depending on the sample, a temporal resolution on the millisecond scale may be needed...plane of excitation light that is thinner than the portion of the sample that is in focus. This technique ...resolution due to the lack of a requirement for high temporal imaging speeds. Standard fluorescence widefield...sectioning, imaging over large distances and high temporal resolution. Traditionally, this type of sample...technique for many of these samples due to its fast temporal resolution and decreased phototoxicity. Lightsheet...

...still many important questions in this exciting field of research. One of the most important: How will ...commute to and invade multiple organs, and most importantly escape the immune mechanisms of the host. The...against cancer. Sipuleucel-T by the Dendreon Corporation is currently the only FDA-approved DC therapy...therapies themselves can be made more effective. Some important factors researchers are beginning to focus on ... of cell types. This cell heterogeneity is an important element of successful tumor progression. A tumor...), and ability to metastasize. There have been reports of clinical trials going amiss and patient deaths...

...nonspecifically soak up your fluorophore package. One important concept in FRET experimental design is that the...donor first where labeling specificity is most important. Donor labeling and the need for specificity Donor...protein molecules contain the donor and, more importantly there is no fluorescence background resulting...in the ribosome, resulting in site-specific incorporation of the attached fluorophore. However, since ...chemical warfare [Vilensky & Redman, 2003]). Most importantly, these compounds cross cell membranes, thus enabling...Ebright, and Richard H. Ebright. "Site-specific incorporation of fluorescent probes into protein: hexahistidine-tag-mediated...protein kinase dynamics in living cells with FRET reporters." Methods 40.3 (2006): 279-286. PubMed PMID: 16908183...

...designated work areas and come with health risks. Importantly, with this method it is not possible to follow...azidohomoalanine (AHA). In the technique, AHA is incorporated into newly synthesized proteins instead of methionine...endogenous protein of interest that also had incorporated the AHA tag after pulse labeling of a determined...FUNCAT-PLA provides both quantitative and spatio-temporal information about the turnover of specific endogenous..."Puromycin inhibition of protein synthesis: incorporation of puromycin into peptide chains." Proceedings...Gabriel. "Inhibition by puromycin of amino acid incorporation into protein." Proceedings of the National Academy...

...changes. In addition to supporting in-person conference attendance, the award now supports virtual conference...research.   Abhi Aggarwal: Glutamate sensing reporters Abhi Aggarwal is a postbaccalaureate research ...third-generation intensiometric glutamate sensing fluorescent reporter (iGluSnFR3) to study glutamate, a signaling molecule...in pMinDisplay (PDGFR anchoring domain), in electroporated in rat hippocampal cultured neurons, imaged...hydropersulfides modulate protein activity and have important roles during oxidative stress. To monitor this...

... an ERK kinase translocation reporter with a mRuby2 fluorescent reporter (ERK KTR-mRuby2). The ERK KTR...control of kinase activity can provide greater spatiotemporal resolution than pharmacological or genetic ...everything downstream of MEK1), bypassing the upstream portion of the pathway and focusing on the downstream steps...protein of interest. If you plan to try this, it’s important to check that the function of the protein and ...Activity with GFP Resources at Addgene.org Find KTR reporter plasmids Find photoactivatable fluorescent proteins...

...want to turn things on or off, permanently or temporarily, we’ve got you covered. Defining your control...requirements To pick the best tool for the job, it’s important to know what the job requirements are. Here are...inducible? How long do you need control? Will temporary control (a few days) be enough, or do you want...than making a knockout and is a great way to temporarily reduce expression of essential proteins (when...expression (more on this below) will both achieve temporary repression of protein expression. Addgene has ...via a custom gRNA, to your gene of interest to temporarily increase expression. There are several flavors...

...workforce working group report. http://acd.od.nih.gov/biomedical_research_wgreport.pdf Resources at Addgene... Training (BEST) awards, which are designed to support new approaches that address the need to train our...Labor Market. On this topic it is clear that many important questions, areas for informed discussion, debate...University of Maryland College Park, is a strong supporter of increasing the number of postdoctoral scientists... collection and analysis. He is also a strong supporter of science Master’s programs. This will be the...

... the known protein concentration. Importantly, you’ll want a portion of your standard curve to plot to...measured using a microplate reader, is directly proportional to the amount of enzyme present in the well ... If your standard curve does not have a linear portion or the OD of your experimental samples has reached...HRP-TMB reaction more quickly. Just make sure to incorporate the dilution factor in your calculation from ... you choose. Once you’ve identified the linear portion of your standard curve, you can calculate the concentration...

...Faculty of Sciences of NOVA University in Lisbon, Portugal. She leads the Glycoimmunology group based at ...glycans. Accordingly, she joined efforts with the Portuguese Association for Congenital Disorders of Glycosylation...PMM2 gene. Videira’s group recently reviewed and reported that PMM2-CDG patients suffer from immunological...wherein it's possible to delete the functional portion of the Nfix gene using Cre recombinase and to generate... construct NFIX expression vectors as well as reporters of NFIX function. They will then mutagenize the... the Glycoimmunology group at UCIBIO, FCT-UNL, Portugal. Her main work focuses on unraveling the pathophysiological...

...delivery include plasmid transfection, RNP electroporation, and viral transduction; but these methods ...Gag protein fused to a Flag-tagged SpCas9 is incorporated into Nanoblades, just like the unmodified MLV...MLV Gag protein. After Gag-Cas9 is incorporated, the MLV protease, supplied as part of the MLV Gag-Pol...into Nanoblades by association with the the Cas9 portion of the Gag-Cas9 fusion protein. A single Nanoblade...gene, while editing was not detected in cells electroporated with RNPs using the same guide. Nanoblades ...influence the level of off-target effects, with electroporation of RNPs generally having lower off-target edits...

...of the yeast TREX-2 mRNA export complex: components and anchoring nucleoporin. Lu et al. Plant J. 2010... (abundant in eukaryotes) A Near-Infrared BiFC Reporter for In Vivo Imaging of Protein-Protein Interactions...green-yellow or orange-red fluorogen A split fluorescent reporter with rapid and reversible complementation. Tebo....  Controls  When designing experiments, it’s important to include proper controls to assess the behavior...target proteins, especially if the termini are important for their interaction. You can read our previous...

... your skills from the start. Just like with any sport, hours of practice are required and no one lab or... grant early! First off, and perhaps the most important bit of advice I can suggest, is to give yourself...Originality is essential for grant success The two most important things reviewers will be looking for in your research...from accounts or HR on the judging panel. It’s important for this person to get a clear and concise explanation...going to be an element of luck involved so it’s important not to throw in the towel if 1 or 2 funding bodies...submission panel. Some funding bodies provide opportunities to attend an overview of what they are looking...

...science advocates is an important way to push the balance from challenge to opportunity. We hope to see you...philosophy of academic science to date. Advocacy opportunities for the future No matter your opinion on these...scientific enterprise will provide challenges and opportunities for us all.  For example, implementation and...challenging. At the same time, there is much opportunity to make headway on long-term efforts to better...we are able to successfully advocate for the opportunities they present. Similarly, ongoing issues that...