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...done using side scatter (SSC) and forward scatter (FSC), which measure light that “scatters” past the cell... the plot that divides it into four, using the location of the cell populations as a guide  (Figure 3....be used for this purpose.  Pro tip! The forward scatter is a proxy for diameter and volume of the cell,...the exclusion of doublets and debris. The side scatter roughly measures granularity, a proxy for nucleic... will have numbers by gated populations; this indicates the percentage of total events contained in the... gated population. Heat maps are often used to indicate the relative density of cells. You can see both...presented. A horizontal line across the peak will indicate the cut-off value for a positive signal (Figure...

...be part of the answer. Problems with written publications Online tools like ResearchGate, GoogleScholar... that the big breakthroughs in my discipline, education, will come from ideas garnered from research in...from the science of others. Improving science communication through video     Video abstracts about research... to be that video is the future in terms of communicating ideas. In 2012, for example, 3 billion hours...scientific videos were for entertainment or basic education (i.e., learn why water freezes) there was a growing...video usurps the written word as the means for communicating research. Short videos of 3 to 5 minutes provide... researchers to leverage the video medium to communicate their research more effectively.” How to share...

...expression vector cannot have p15A origin of replication F- ompT lon hsdSB (rB- mB-) gal dcm (DE3) pLysS...expression vector cannot have p15A origin of replication F- ompT lon hsdSB (rB- mB-) gal dcm (DE3) pLysE...Expression vector cannot have p15A origin of replication F- ompT hsdSB(rB- mB-) gal dcm (DE3) pRARE2 (...Expression vector cannot have p15A origin of replication fhuA2 [lon] ompT gal (λ DE3) [dcm] ∆hsdS/ pLemo...Expression vector cannot have p15A origin of replication F-, Φ80ΔlacM15, thi, lac-, mtl-, recA+, KmR Expression... commercial strains carry the DE3 lysogen, as indicated by the name of the strain. Conversely, other strains... for positive selection and a p15A origin of replication, making it incompatible with other p15A plasmids...

...Addgene ID number, your publication will be listed on our website as a “Publication Citing this Plasmid” ...upon other researchers being able to find their publications in the first place. As a curator of the thousands...make it easier for you to find the plasmids and publications relevant to your research interests. We do this... this by promoting citation, supporting sophisticated search, and curating plasmid collections. Addgene's...number as well as the reference for its original publication. To help facilitate this process, we explain ...fruits of your labor and won’t have to repeat the modification. This will save both time and money. A quick...whether the plasmid has been used in your system. Sophisticated search You can search for specific plasmids ...

... use the GAL4-UAS system along with precisely truncated attP sites to control recombination of the gene...of PhiC31. In the SPARC system, attP sites are located on either side of a stop cassette, upstream of ...amount of labeling, you can alter the attP site. Truncating the attP site upstream of the stop cassette results...population is directly correlated to the length of the truncated attP site. The three SPARC variants, based on ...intermediate (d), and sparse (e) labeling of the calcium indicator, GCaMP6f, in the Drosophila optic lobe. Image ...Image from Isaacman-Beck et al., 2019. Applications for SPARC SPARC technology is versatile and can be ...dendrites of T5 neurons by placing the calcium indicator GCaMP6f in the effector position, as well as to...

...study its function is one of the most popular applications of fluorescent proteins. These fusion proteins...steric hindrance. For instance, GFP may occlude catalytic sites or obstruct binding sites or motifs that...that are necessary for post-translational modifications. Despite these potential issues, GFP has been used...linker sequence short In the early days of GFP applications, many were concerned with steric hindrance by...N-terminus of the donor fluorescent protein can be truncated to increase the FRET efficiency. Creating fusions...fluorescent protein and protein of interest (POI). To indicate the sequence, people often use terminology like...  Determining other aspects of functionality (catalytic activity or interactions) can be challenging. ...

...well as protease sites to facilitate protein purification. We hope this kit will allow you to quickly ...for a variety of synthetic biology or protein purification studies in E. coli. 800 and 1900 Series If you...polycistronic E. coli expression constructs. Some applications of plasmids from this kit include the production...and Charles E. Melançon III. "Detection and Quantification of Ribosome Inhibition by Aminoglycoside Antibiotics...designed the pCri system to simplify the protein purification process by making it easy for a researcher to... blog post on the system Easing the Protein Purification Process with pCri Cell-free protein expression...Daniel, et al. "A pull-down procedure for the identification of unknown GEFs for small GTPases." Small GTPases...

...most inclusive databases of post-translational modifications, identifies a staggering ~250,000 phosphorylation...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...codons (TAG), replacing them with TAA (7). This modification allowed the deletion of release factor 1, the...optimize protein production using standard protein purification strategies; expression should be optimized for...-incorporated to non-phosphorylated protein. Verification of site-specific pSer incorporation can also...programmable protein phosphorylation." Nature communications 6 (2015). PubMed PMID: 26350500. PubMed Central...

...Scientific Publication, where diverse approaches to peer review were discussed. Science communication opportunities...developmentally programmed regulation of DNA replication and gene transcription during vertebrate embryogenesis...There is a rich and active community of science communication professionals who are thinking about, for example...access affects financial models in information publication. Take a look at the blog ‘Scholarly Kitchen’ ...opportunities If you are interested in the science communications world, you might consider an internship or ... is enjoys broad engagement through science communication with a variety of audiences. She focuses at ... science communication...

... This allows them to replicate viral plasmids with the SV40 origin of replication. To generate a virus...isolating the cell line all the way to modern applications. What the HEK is it?   HEK cells were first...cell biology, there are thankfully many other applications that they are useful for! What the HEK is it...cell line and we will highlight some of those applications here.   Virus production HEKs are the go-to ...proteins at high levels. In a case where clinical applications of recombinant proteins are necessary, HEKs ...significantly reduces the cost of production. Research applications There are two areas of research where HEK cells...vulnerable to viral infection. Yes, your cells can catch a cold—or worse! HEK-293T and its derivatives are...

...  After identification of ABE activity, XMAS-TREE can also be used to purify these modifications and work...ABE-targeted cells By Ellie Adams Single base modifications through ABE or CBE's have the potential ability...point mutations. They allow single-nucleotide modifications to edit DNA without requiring a double-stranded...difficult to modify in the past.   Figure 1: Identification and enrichment use of XMAS-TREE. a) Cells are...mCherry and GFP expression to check for ABE’s. b) Quantification of base-editing efficiency using XMAS-TREE-...methods at various genomic loci in four different categories.     Brookhouser, N., et al. BMC Biol 18...MYC-dependent cancer cell lines. It can be applied for identification of essential MYC binding sites in a wide range...

...protein-only transduction method and its potential applications to CRISPR/Cas9 genome editing.   Protein-only...the HIV protein Tat. Although CPP fusions can translocate a cell membrane, the presence of the CPP may ...that components in the buffer used for protein purification must be helping the untagged protein enter cells...Diego D'Astolfo. CRISPR/Cas9: a potential application of iTOP Having tested iTOP using Cre-lox recombination...showed up to 70% resistance to diphtheria toxin, indicating a 70% frequency of biallelic mutations. Importantly...Importantly, hESCs retained their stemness, indicating that iTOP may be valuable for CRISPR/Cas9 stem cell...potential for off-target effects. Third, protein purification (once a protocol has been established) may be...

...Mechanism of DNA replication fork breakage and PARP1 hyperactivation during replication catastrophe. Science ...availability alone, indicating an intrinsic shift in enzyme processivity. Structural data implicate specific residues...ABE8e*A showing dimerized TadA subunits, with the catalytic domain in dark red and the docking domain in pink...(UC Irvine) showed that APOBEC3B activity at replication forks induces APE1-dependent abasic site cleavage...as both a prognostic marker and a potential stratification marker in treatment decisions. Shelby Devenport... only a few years ago. The gap between first publication and widespread adoption has never been shorter...fundamental mechanistic insights to translational applications. If you presented at Deaminet 2026 and have...

...Gibco organizes its app into four categories: tools, analytics, education and data manager. The tools are...are five FREE science lab apps that have been educational and great for lab life. These apps prevent you...lab notebook. Lab Archives is a multipurpose application that allows you to share your data with colleagues...other nice features. This app also has a great educational section. This section holds the Gibco Cell culture...directions including which buffers to use and that indicates if a sequential digest is needed. The enzyme finder... to this app; the user can search by sequence, category of enzyme, or name to discover all the information... Addgene.org Find Protoctols for Your Research Catch Up On Your Molecular Biology Basics ...

...together a grant application? Are you familiar with the publishing or patent application processes? How ... pages in a Curricula Vitae to showcase your publications, academic performances, and research experiences...information. Think "skills," not just "research and publications” What are the skills you developed through your...and writing was how I got my current job as a Communications Coordinator. Wondering how you can develop ...So You Think You have Skills.   Get Science Communication Practice with a Student Group Like Science in...Blogger! Theresa Liao (@theresaliao) is the Communications Coordinator for the Department of Physics &... a research career, she transitioned into a communications role instead after she completed her MSc in...

...your application a whole week before the deadline. The day of submission, review your application with... their dream.  To help students focus on the application process, and simplify access to academic resources...Scrutinize every opportunity prior to starting an application to assess your eligibility, interest, and career... detailed. Beyond the resume, a cover letter communicates your interest in a specific program, and institution...February represent peak season for internship applications. Thus these noble individuals are inundated ... with a stellar letter of support.        5. Application submission  Set a calendar reminder for every...about outreach among low-income communities to educate parents about resources that can help their children...

...discuss automated protocols. 12:00 PM--Catch up. Finish any communications with my UK team members before they...getting to talk to so many different people! I communicate with customers, software developers, as well ...from interacting with others, and have strong communication skills to tailor your message to any type of... to solve is clear, but the solution is more complicated. You’ll need to be creative to be able to work...team in the UK is five hours ahead, so I’m always catching up in the morning! 9:00 AM--Technical implementation...out. 4:00 PM--Respond to customer help tickets. Catch up on customer help tickets, and respond when I ...technology, software, and science. She loves to advocate for the customer and help build software and technology...

...ITRs serve as a type of origin of replication to initiate replication of the dsDNA virus. Viral packaging...plasmid while the other plasmids with packaging or replication components are called helper plasmids. Below,... production, they can be helpful for certain applications and aid in ease of use. Our lentiviral guide...AAV are rep and cap. Rep is required for viral replication and also aids in infrequent genome integration...a helper plasmid and mediate efficient viral replication. More information on these viral genes is available...particles. E2A/E2B are responsible for viral replication, with E2B being the primary viral polymerase....is missing the endogenous E1 gene, making it replication deficient. This is typically supplied by the ...

... its function, mimic a biologically relevant modification, or add in fluorescent or chemically reactive...function relationships or, in more practical applications, allow a protein to be more effectively used... you do to any number of experiments with. The catch The rapid expansion of GCE led to the creation of... of many new tools and potential applications. But this focus on development, along with the inherent ...’t give it a try. Optimization Why is GCE so complicated? Well, while the techniques are straightforward...ligation through introducing ncAAs with protein modifications of interest or any of a huge variety of ncAAs...has “10x the impact of PCR. But it’s 100x more complicated.” And Ryan Mehl would know. He’s the director...

...Our educational content  Have you ever wondered how we choose topics for the Addgene blog? Or why we...everything else we do! The goal of Addgene’s educational content Addgene truly believes that sharing speeds...antibody and content team, we identified what applications should be included and what basic information... The guide was then assigned to our science communication specialist, Rachel, to write. Identify expertise...through an editing process and final check before publication.   Do research! Nearly everything we write entails...Guide figures. This requires a fair amount of communication between our content team and our graphic designer...graphic designer upon request that the science communicator "just sketch a western blot out quickly." Final...