...different questions than those on the prep calls, but often the prep calls provide good ideas for interesting ...possibly with an organizer) should set up a 30 minute call with each panelist to get to know their style and...better at remembering these things on a relaxed phone call than when first confronted on a stage in front of...long introductions. The moderator asks questions, calling on 1-2 panelist for each question in a way that...

...the right fit for you without the financial and logistical stress that accompanies traveling for campus ...Visits After receiving applications, universities typically invite a select group of students to continue ...and each other. Before COVID-19, financial and logistical obstacles could make touring potential programs...in-person interviews for a variety of reasons (logistical, financial, etc…) now have options to fully engage...format of interview from a travel day to an online call in response to challenges on their end. Though the...

... more than 10 modules often rely on two-step hierarchical approaches using different Type IIS restriction...stability, and buffer compatibility have placed practical limits on single- or two-step assemblies. We have... While T4 DNA Ligase, the mainstay of most biotechnological cloning efforts for over 50 years, prefers... plate Fidelity of assembly (percent correct) Calculated colony totals Per 2 μl assembly reaction Per...outgrowth volumes described in Figure 3, with calculated yields from entire outgrowth built from 2 µl ...

... molecular biology-grade agarose containing physiological concentrations of salts and glucose. If we circled...fluorescent protein), fluorescence microscopy can theoretically be used to select specific wells from the 48... terms of growth rates, morphology or other physiological properties. An advantage of this technique is...properties, either initially, or using secondary biological screens. For example, for cancer researchers,...Neurobiology and Anatomy at the University of Maryland Medical School in Baltimore. She is particularly interested...

...BV510 and AmCyan have almost identical emission spectra but may be called BV510 on one cytometer and AmCyan...for one another. This overlap in spectra is often called "spillover." For example, if your flow cytometer...shift towards the X axis to become a perpendicular vertical line.     In general, most spillover between samples...research technician in Arlene Sharpe's lab at Harvard Medical School.   More resources on the Addgene blog Introduction...

...Preger et al., 2020. Preger et al., Journal of Biological Chemistry 2020. https://doi.org/10.1074/jbc.RA120.01289...research, which required protein compartments to co-localize multiple enzymes in a reaction pathway, the MPyV...expression selectively in Parvalbumin containing cortical interneurons. We now provide ready-to-use AAV ...vectors expressing GCaMP6f with this promoter. Image calcium transients in PV interneurons with AAV-S5E2-GCaMP6f...terminal providing better signal to noise for imaging calcium dynamics in fine structures. We have expanded our...

...introducing the delivery method in 2014, Manuel Goncalves lab report that AdV-mediated transduction of gRNA...Gonçalves and his colleagues at Leiden University Medical Center are now available at Addgene along with ...and very efficient introduction of DNA into therapeutically relevant, non-transformed mammalian cells. ...and used AdV vectors to edit genes in several biological systems. It has been used for instance in inherited...Viral vectors for gene therapy: Current state and clinical perspectives. Biochemistry Moscow 81:700–708 ....

...in-house protocol. DNA yields for this procedure are typically in excess of 100 ng/ul, more than enough DNA to...be viewed more as a guideline, since they are typically based on the copy number of the empty plasmid ...improved plasmid DNA yield. Culture conditions play a critical role in plasmid DNA yield. Different plasmids ...are generally, but not perfectly, correlated. Typical growth times for high copy plasmids in standard...miniprepping at various time points. Measuring the optical density of the culture can indicate the number ...

... and ways to navigate it, we’ll search for the calcium biosensor, GCaMP, and click through to the plasmid...distribution. If your experiment involves delivering a calcium biosensor using AAV, you might decide you’d really...? The available filtering options now change dynamically based on the type of material and specific search..., followed by a convenient table of plasmids specifically designed for this purpose: Still stumped? Ask...results at all, let us know! Contact our scientific technical support at [email protected] and let us help.  ...

... of these techniques have in common? They all typically require secondary antibodies, frequently of the...are derived from an unusual type of IgG antibody called a heavy chain antibody (HCab), which are unique...different species, nanobodies are useful for colocalization labeling of up to 3 targets (Pleiner et al ...desired fluorophores. In figure 4B below, triple colocalization is possible even when staining cells with three...multi-color staining workflow yields similar localization patterns as cells stained for one target (compare...

...Ample distribution channels both digitally and physically. Easy to incorporate visuals if needed. Traditional...means of scicomm. Digital or print distribution.  Typically limited to a single page of information. Primarily...levels and interaction. Can bring in things to be physically interacted with. Public speaking is a common ...internet access. High probability of real-time technical difficulties Exhibits Medium to high Audience...exhibit). No remote access; limited occupancy. Typically needs to be staffed. Mixed Media Medium to...

... science teachers at our school put out the call to local parents who would feel comfortable speaking ...parents who can answer some of the surprisingly technical questions elementary students can ask. It doesn... dangers of people using the technology in an unethical manner, but I also tried not to be an alarmist...fields in biology, are moving so quickly that it’s basically impossible for textbooks to keep up. Even the ...

...especially in the sciences and by extension to scientifically oriented nonprofits. Like Deb, I worked for...available through not-for-profits, universities, local and national government offices, schools, healthcare... arts? Do you find satisfaction in helping your local food shelf raise money? Do you have experience with... as a grant writer and/or grant reviewer with a local nonprofit that aligns with your passion to get grant...Join the Grant Professionals Association (GPA)  local GPA Chapters, and the National Grants Management...

...molecules, forming a “handcuff” structure that physically blocks replication machinery when plasmid copy...It is also worth noting that plasmids can be symmetrically incompatible, meaning that both plasmids are... from the cell with the same probability, or asymmetrical, where one plasmid lost from the cell at a higher...deal with the more sinister plasmids of the microbiological world. (Want to know more about these plasmid...high copy incompatible plasmids that lead to asymmetrical plasmid loss of the usually large, low copy ...

...post was updated on Jul 27, 2020. CRISPR, and specifically Cas9 from S. pyogenes (SpCas9), is truly an ...specified by the crRNA sequence, it adopts an enzymatically “active” state rather than reverting to an inactive...propensity of Cas13a to bind and cleave RNA non-specifically. The ability of dCas13a to bind target RNA without...modulating splicing, and regulating expression of specifically targeted RNAs. Finally (and perhaps most interesting...observed shRNA off-targets. They subsequently used catalytically inactive dLwaCas13a as a programmable RNA-binding...

...the Janelia Research Campus of the Howard Hughes Medical Institute. Are you currently on or planning to ...rejection of a model and no clear alternatives. Typically, a research vision is broken up into three aims... basis of a single grant. Either way, think realistically about the time and resources required to feasibly... aims in more depth in the remaining time. The typical chalk talk structure is: What is the big question...the Janelia Research Campus of the Howard Hughes Medical Institute. His interests include the quality control...

... this post I’d like to delve deeper and look specifically at the characteristics and capabilities an ELN...facilitate true reproducibility. The following are critical: (1) connectivity to other data sources used in...likely stored in institutional and lab servers specifically designed to hold large amounts of data. Although...group’s research – it makes no sense and is not practical to move all this data in bulk into a lightweight...lightweight tool like an ELN. It is nevertheless critically important to be able to conveniently reference specific...

...product purity of 66%. Pro tip! Abasic sites are also called apurinic or apyrimidinic sites and abbreviated ...are the same protein!) (Chen et al., 2024). They called their initial editor AXBE, for A → X base editor...in a G → Y (Y = T or C) base editor, which they called glycosylase-based guanine base editor (gGBE) (Tong... Therapeutics team has deposited their editors, called gTBE and gCBE, with Addgene. gTBE can introduce...site A DNA site with the nucleobase removed. Also called an apurinic / apyrimidinic (AP) site. UNG / UDG...

...In Addgene’s early days, we would ‘spot-check’ critical regions of plasmids deposited with us via Sanger...development teams, frequent updates allowed us to scale our lab work as we grew. As the number of deposited...us to switch between executing our pipeline on a local computer or in a cloud environment. Given the throughput...jumping back and forth between running the pipeline locally and in the cloud. Creating a bioinformatics workflow...script file (containing our new process) on our local computer. Eventually, our workflow would handle ...

...experimental design and its potential for clinical applications. Specifically, efficacy rates of Cas7-11 were reported...current knowledge. The Abudayyeh-Gootenberg lab (also called the AbuGoot lab), a joint lab run by Omar Abudayyeh...originally suspected that this new Cas7-11 might be phylogenetically classified as the more complex Class I systems...switched up the serine integrase for one which can enzymatically take a genetic sequence and integrate it into...