...next logical step after technical mastery. However, being a great scientist doesn’t automatically translate...cohesive team environment. Neglecting communication skillsClear communication is the backbone of effective management...transition While a PhD equips you with exceptional technical skills, transitioning into management often means...stepping into unfamiliar territory. Unlike solving technical problems in the lab, management involves a different...time, I discovered that I not only enjoyed the technical aspects of my work but also found fulfilment in...your team's—can lead to frustration. Clinging to technical tasksDelegation can feel unnatural for those accustomed...myself to focus on my core responsibilities, which typically revolved around the broader vision rather than...

...weather befitting of the ominous political climate in which American scientists find themselves today,...we spend years being taught to communicate in a very specific, technical manner, so much so that we often...also a science communicator, UBC Public Scholar, and a member of the Vancouver SciCATs. Follow her on ...March for Science from researchers all over North America. I hope that these perspectives, the optimism they...Alina Chan is a postdoctoral researcher at Harvard Medical School working on human artificial chromosomes....election.   Even as we marched in solidarity with our American neighbours, the fight is far from over here at...April because devaluing science has alarming implications for our public policies—many of which impact...

...your specific application. Consider also that there may be variations within an application. Take immunohistochemistry...website highlights a variation of your application, a publication successfully used it in a similar way...exciting new project aimed at understanding how a critical cellular pathway is regulated. You’ve read all...choosing an appropriate antibody first look at the application-specific data provided on the manufacturer’s ... a linear antigen tend to work very well for applications that denature a protein to its primary (linear...the most similar to yours. Take a look at the publications cited on the manufacturer’s website, which may...plan to. To secondary or not to secondary?  The application will also determine whether you should use an...

...regular part of basic biological research as well as clinical therapy. These biological tools are useful because...being engineered as biological tools, thought was given on how to keep the replication of virus and its ...the component that physically integrates into the host genome, the virus is replication incompetent, meaning...envelope plasmid), and the polymerase (Pol) for replication and Gag structural protein on yet a third plasmid...meaning it can’t replicate and continue the infection cycle once inside the host. While the virus can still... this safety feature prevents the virus from replicating and spreading in non-target cells, including ....    Viral cargo-specific safety   After replication competency, the second concern with virus safety...

...Finding an antibody that works for your application, specifically detects your protein, is species compatible...(stay tuned for our affinity purification tag blog!).   Applications of epitope tags   The...protein purification is not ideal with Myc, as low pH conditions are required during the purification, which...and have been widely used across species and applications. In this blog, we will review the most common...that are bound by antibodies for the purpose of purification or detection of the tagged protein. These peptides...not exist, or they don’t exist for a specific application, or they exist but are unreliable for a variety...almost certainly already been optimized for your application. Depending on the tag, directly imaging or tracking...

...of its biggest limitations is that sequence modifications can only be made at restriction enzyme cut sites...homologous recombination. It allows for direct modification of DNA within E. coli and is independent of ...engineering.  It can be used to make an assortment of modifications: insertion and deletion of selectable and non-selectable...however, only Beta is required when generating a modification with an ssDNA substrate.  Gam: Gam prevents ...insert site (Figure 2, top).  These primers are typically ~70 nucleotides long (20 nucleotides that anneal...regions flanking the target site).  The best applications for dsDNA inserts include: large insertions ...fragments, such as gene replacements and tags.  The typical frequency of recombinants is 1 positive clone out...

...2012) scFv Antibody: Principles and Clinical Application. Clinical and Developmental Immunology 2012:1...therapeutic applications where longer retention times often increase therapeutic efficacy (Ahmed et al...scFvs are often genetically fused to other proteins, which has led to their application for basic and translational...cheaper to generate since it’s genetically encoded.   Translational applications of scFvs: Just like antibodies...blood clearance, which is useful for imaging applications; better tissue penetration which is useful for... for therapeutic and imaging applications; and reduced immunogenicity when administered in vivo due to... translational research.  Basic research applications of scFvs: The SunTag system uses an scFv to amplify...

.... Lajoie, Marc J., et al. "Genomically recoded organisms expand biological functions." Science 342.6156...most inclusive databases of post-translational modifications, identifies a staggering ~250,000 phosphorylation...phosphorylation events on the activity of a given protein? Classical methods for studying protein phosphorylation A...steric and charge-based nature of the phosphoryl-modification; they are both much smaller, and carry only ...thus cannot reflect the true state of protein modification within the context of its appropriate regulation...plasmid. This includes 4 copies of a tRNA that specifically recognizes amber stop codons (TGA) to encode...effects on cell growth. The C321.ΔA strain was genomically recoded to eliminate all 321 amber stop codons...

...ASAP5, a faster and more responsive genetically-encoded voltage indicator (GEVI) than previous versions. ASAP5...responsivity in screening ASAP3 variants. B) Electrical and optical recordings of spontaneous activity in human...protein-coupled receptors (GPCRs) are critical components of many physiological pathways and represent the largest...by endogenous GPCR. Example applications include large-scale pharmacological profiling or interrogation...single-molecule biophysics Fast and responsive voltage indicator ONE-GO biosensors kit Antibodies for neurodegeneration... an enhanced pU6cm promoter, viral replicon amplification of cargo, and heat treatment.  Figure...Arabidopsis, suggesting the improvements may be broadly applicable to dicot species. In addition, they generated...

...Foundation, an organization working to advance biological engineering openly so as to benefit all people...Initiative. Endy's team made news in March with a publication in Science describing the development of “transcriptors...with the BioBricks Foundation to distribute the physical DNA samples, for when that's the preferred mode...parts must flow.' People must be able to use genetically encoded functions - the words in our language...operating system for engineering an entire cell. Typically when people use plasmids, they are trying to do...the entirety of a cell. That's a big long-term technical challenge. We have a strategy that organizes how...work, and it begins with the central dogma - replication of DNA, transcription of DNA to make RNA, translation...

... ITRs serve as the origin of replication and are required for replication, packaging, and vector persistence...viral rep (replication) and cap (capsid) proteins, which bind to the ITRs and initiate replication. ITRs are...important elements of AAV and rAAV vectors, modifications can drastically affect gene transfer efficiency and ...Adeno-associated viruses (AAV) are an important biological tool that allow scientists to express a gene ... with biorender.com. ITR structure  ITRs are symmetrical 145-nucleotide sequences that flank the ends ...binding site for Rep78 and Rep68 to initiate replication and a terminal resolution site that serves as...are noted. Created with biorender.com. ITR modifications and types  Since ITRs are one of the most important...

...COVID-19 cases each week. Statistical Modeling Note -- If statistical modeling isn’t up your alley,...breaking the statistical independence assumption of the Poisson process. This modification also eases the...learning and statistical methods for fundamental scientific discovery and high-impact applications.       Alvin... of Chemistry and Chemical Biology at Harvard University; and Howard Hughes Medical Institute Investigator...months, pausing or terminating employment for a significant fraction of the workforce. Wet lab scientists...subfields.  Plasmid requests correlate with paper publication The dataset includes 11,426 plasmid requests ...14%). Spikes in plasmid requests correspond to publications of popular papers. Figure 1: Plasmid requests...

... be run in duplicate or triplicate. The closeness of the data points generated by replicate values provides... the standard dilution series in duplicate or triplicate. Replicates allows you to calculate the curve...blot. Standard curves are critical for a number of popular scientific applications, and the quality of a ... deviation of each replicate measurement divided by the mean of all the replicates multiplied by 100. ...regularly, and mix samples very well.  Use duplicates or triplicates to increase the accuracy of your experiment... value. Though it varies from assay to assay, typically the experimental value should be within 15% of...correlation between the actual values and the predicate values and measures how well the data points fit...

...caudal NST. (A-C) Low-magnification images showing viral spread. (D-F)  High-magnification images immuno-stained...vector performance. Yet, most of the technical data needed to replicate and build on others’ results is available...Produce Chemically and Functionally Distinct Vectors. Molecular Therapy - Methods and Clinical Development...and community concerns AAV vectors are complex biological reagents; many factors, like storage buffers ... right AAV for your system or to successfully replicate published works. Of particular importance to these...improve our Addgene Data Hub, 76% of respondents indicated they wanted to see more negative data (Figure ...out of fear it points to flaws or gaps in their technical ability. But, especially for AAV data, that may...

...and often cannot eradicate metastasis or eliminate the probability of a relapse. Chemical and radiation ...the tumor, this results in frequent physiological and morphological changes. Tumors differ in their genotypic...-level barricades put up by cancer. As mentioned in this article, cancers differ in practically every ...immune response to the development of tumors is a complicated and dynamic phenomenon that is interestingly ...either due to age, infections, or other strenuous biological changes. In contrast to the acute immune responses...themselves from attack. It is a region of incessant chemical activity, and consists of other cell types, especially...attack the cancer cells are also suppressed by chemical signals such as IL-8, IFNγ, and colony stimulating...

..., can be expressed from lentiCas9-Blast. Purpose of LentiCas9-blast LentiCas9-blast is a Cas9 expression...single-vector systems typically express Cas9 and a gRNA from the same plasmid. LentiCas9-blast virus can be...began with only a few inventory items offered domestically, but by the end of 2016, we expanded our viral...2016 were (drumroll please)... Top Lentivirus: lentiCas9-Blast Most of Addgene’s lentiviral inventory is... preps. Cas9 is the headliner of most CRISPR applications and can be expressed from your cell line of ...2014). It is a dual-vector system, meaning it is typically used in conjunction with a second vector that ...several benefits. First, the low viral titers typically associated with Cas9-expressing lentiviruses will...

...By Alyssa Neuhaus Genetic modification of microglial immune cells is critical for central nervous system...mice. B) Quantifications showing high microglia specificity of GFP expression for the indicated ssAAV5s...ssAAV5s. C) Quantifications showing high microglia transduction efficiency for the indicated ssAAV5s. Figure...Emily Bentley Building on their CROPseq method for optical pooled CRISPR screening, Paul Blainey’s lab has...compatible with enrichment, single-cell sequencing, and optical pooled screens in human cells. Addgene offers vectors...3’ long terminal repeats (LTR). The 3’ LTR is duplicated during lentiviral integration, producing a second...described in Anzalone et al., 2019, including modifications to the Cas9 or reverse transcriptase enzymes...

... University where she engineers genetically encoded voltage indicators. Tian developed a video-based high-throughput...red‑shifted archaerhodopsin‑derived genetically encoded voltage indicator which had improved performance in...the improved voltage indicator coming from the screen to understand mouse cortical and hippocampal neural... Haoyi Qiu He Tian Every two years, we seek applications for awards to early career scientists using ...virtual conference attendance. We also opened up applications to research technicians and undergraduate researchers...the brain. “We hope that iGluSnFR3 will enable significant steps forward in our understanding of synaptic...the chromophore that are susceptible to redox modification, the MPST hydrosulfide can be transferred to...

...in PubMed and choose the appropriate publication. This publication will be listed at the top of the final... "Published plasmids" or "Unpublished or pre-publication plasmids" and set a desired distribution status...quickly associate your plasmids with the original publication where you described them. Search for your article...plasmids. You can submit "Unpublished or pre-publication plasmids": Unpublished plasmids or plasmids that...accepted. Addgene also encourages submitting pre-publication plasmids so they can be available when the paper...preprint submission, we are happy to update the publication information once the manuscript is published ...your desired distribution status: Hold for publication: we will initiate our MTA process and perform...

...likely that the shifts caused to the practical and philosophical landscape of the scientific enterprise...Director of Media Communications, Harvard University Ray Howell -Founder of Howell Communications Philip Brenner.... Revitalizing biomedical research: recommendations from the Future of Research Chicago Symposium F1000Research...and keynote by former congressman and CEO of the American Association for the Advancement of Science, Rush...solutions. The event also culminated in a joint American Society for Biochemistry and Molecular Biology...union dues. Time will tell the extent of the implications of this ruling and whether graduate students...focus toward advocacy and action with sessions specifically chosen to better define productive routes for...