...education? Let us know about your educational events in the comments section below! Additional Resources Visit...Last week, 20 students and four chaperones from Denis-Yarmouth Regional High School visited our labs ...Learning about careers at Addgene Afterwards, the students split into two groups for an in-depth look at ...Biology Lianna Swanson was very popular, with the students excited to see a real working lab full of busy...Operations Coordinator: “Kory was awesome!”. The students also got to learn about intellectual property ... technology was a rousing success! Many of the students were interested to see so many different career...such a great day, we look forward to hosting more students next year! Does your lab or organization do work...

...majority of the respondents (70%) used a lipofectamine-based method to deliver NgAgo ingredients into the cell... questions about the NgAgo protocols used by respondents (transfection method, amount of each component...successful indel formation at a target site. Also, 41 respondents attempted to use the technique for epitope tagging...Survey The take-home message While most of the respondents have been struggling to achieve genome editing...generation and epitope knockin was achieved by a few respondents suggesting that NgAgo can indeed be optimized... take-home message thus far, based on survey respondents "Final Verdict", is that more time is required...Addgene Blog - Leave suggestions and tips in the comments section below or contact [email protected] if you'd...

...Spiking your media with supplements such as magnesium salts, buffering agents, and/or supplying additional...Use a strain optimized for plasmids with repeat elements, such as viral plasmids with inverted terminal... and prep high quality DNA that contains these elements. Be aware of endA+ strains Strains like Stbl3 ...including HB101 and derivatives, release large amounts of carbohydrates that can inhibit the cell lysis...of protein, not plasmid DNA. If you need large amounts of DNA from a plasmid in a protein expression strain...culture media Changing to a richer media or adding supplements can improve your cell density, leading to an ...normally recommended antibiotic concentration. Add supplements to the media You can achieve higher cell density...

...phenotype can lead to a wealth of additional experiments investigating the role of individual genes or...carry out so-called “forward genetic screening experiments”, it is certainly the most robust. In this blog... genome-wide screens and highlight some of the reagents that have been made publicly available through... and versatility. CRISPR/Cas9 consists of two components: the non-specific endonuclease (Cas9) and a single...72005145@N00/5600978712/ What CRISPR-based reagents are available for genome-wide screens? The goal...gRNA-containing plasmids. CRISPR genome wide-screening experiments commonly use lentivirus to deliver a pooled population... mind that libraries are only as good as the experiments you use them for! A well-developed biological...

...scheduling was in part defined by both the number of participants and the standard academic calendar. So you want... their schedules and read the paper (PS: All participants should read the paper ahead of time!). This ...rather than diving deep into the individual data points. All formats should be an open forum where people...figure) if the primary purpose is training graduate students in critical reading. If the aim of the journal...and other subsequent meetings. The number of participants, breadth of the discussed topic, and level of...has lost both its focus and critical mass of participants. This is especially true of journal clubs where...projects have taken on a larger priority for the participants. Perhaps people feel confident in their own ...

...of an objective you have, you can’t resolve two points within ~250 nm of each other. For reference, individual...most SRM techniques also require specialized instruments and technical expertise, limiting their accessibility...relative positions, allowing you to resolve individual points with a conventional, diffraction-limited light ..., ExM protocols utilize relatively inexpensive reagents and are readily adoptable by labs already familiar...will link biomolecules to the gel and add gel components to polymerize. Step 3: digest the sample to allow... “super-resolution” technique that uses common reagents, straightforward protocols, and standard microscopes...isotropic expansion depends on how well the gel components penetrate the sample, how well the gel polymerizes...

...dyes. Acknowledgements I thank Brett Mensh, a wordsmith soldier of fortune, for helpful comments. Many...reverting” to small molecule dyes seems daunting—no one wants to spend their days microinjecting fluorescent conjugates...easier and more functional in complex biological environments such as live cells and tissues (Figure 1). These...activating proteins (FAPs) – These modified antibody fragments bind and enhance small molecule fluorogens (Figure... cell permeability. In past decades, further refinements to this established dye structure have yielded...living cells, especially for advanced imaging experiments. We continue to develop derivatives that have...for many (if not most) fluorescence microscopy experiments. However, when your budget gets a bit thin, small-molecule...

...proteins are provided in trans (see Other Important Components of EBOV Minigenome System). However, minigenomes...RNA viruses (reviewed in 2, 3). Other important components of the Zaire ebola virus minigenome system Similar...plasmid with all four genes and necessary control elements for each would be relatively large. Cloning the...useful research tools. In addition to the reduced requirements for biocontainment precautions, these systems... can be examined independently. In addition, experiments can be performed in the absence of some viral...efficiently transfected into cells. All of these components are necessary to efficiently reconstitute a functional...cell lines that are highly transfectable.  The amounts of each plasmid transfected into cells are also...

...way Fun events don't have to cost a lot, or anything at all really! We've had plenty of events that included...throw in some passion for our mission to share reagents with scientists around the globe, the end result...effort to continue the tradition and plan enjoyable events for employees. We now have a team of three (myself...myself included) that plan big company events and fun things to do throughout the month. Although we are...building activities a priority and have organized many events over the years. To keep the organizational side...Tips! 1. Take suggestions Some of Addgene’s best events have been at the suggestion of an employee, or ...outside of your ordinary environment Team building events like Escape the Room or Habitat for Humanity take...

...designed 2 variants of EosFP that can withstand these conditions- mEos4a and mEos4b (12). Both variants are ... random mutagenesis screening for enhanced RFP variants. PA-mCherry (E26V / A58T / K69N / L84F / N99K ...respectively. When illuminated with violet light, both variants are photoconverted to forms that emit brighter...temperature below 30°C and this is not ideal for experiments in mammalian cells. Dendra2 is another monomeric...PA-FPs. Several other irreversible PA-FP spectral variants have been discovered such as Kaede and PS-CFP2...protein for your experiment Similar to performing experiments with standard FPs, before choosing a PA-FP for...even begins! If you are performing multicolor experiments, you will want to be sure that the light used...

..., including investigations into environmental pollutants and health, embryo growth, brain function, and...visualize expression data in useful ways. Zebrafish mutants In the 1990’s, two revolutionary large-scale random... mutagenesis screens produced ~1,500 zebrafish mutants that are still used in labs across the globe (Nüsslein-Volhard... in congenital heart defects and cancer. These mutants are used extensively to study development and disease... Database ZFIN actively curates information on mutants and other resources. Targeted gene deactivation...deactivate genes without spending months generating mutants. Unexpectedly, phenotypes sometimes differ depending...embryogenesis live. The origins, divisions, and movements of each of the thousands of cells in the developing...

...processes enabling organisms to control their movements and behavior. These functions and others are controlled..., that facilitate immunohistochemistry (IHC) experiments with tag-specific antibodies. The most famous... cannot be detected directly in live imaging experiments. It is therefore difficult to use epitope tags...fluorescent and can therefore be used in live imaging experiments crucial to studying neuronal circuits and dynamics...protein localization, isolation, and tracking experiments (Rizzo, 2009). With many different FPs that emit...researchers can also use multiple FPs in single experiments and visualize numerous molecules or structures...only a single channel. In multi-color imaging experiments, the second color is usually provided by red ...

...researchers and PhD students. Everybody in science knows that there are many ways your experiments can go wrong...researcher doing some experiments in Brazil can have the same issues as one doing experiments in France. We can...samples, or a dysregulated water bath boiling your experiments, just about anything can disrupt your bench work...research. Find Protocols Lab Tips to Keep Your Experiments Running Smoothly and "Gnome-Proof" This is why...laboratory has never faced an odd situation with experiments? Our art simply mirrors our lives at this point...discovered that it can be good for you and your experiments to think outside the box, be creative, and put...

...on during the symposium was “How many graduate students/post-docs?” You can find some of the more detailed...even than the $21/h [on the low end] that grad students are paid before fringe benefits); what they don...receive as postdocs is not transferrable to other environments. Slide from Paula Stephan’s Talk (All slides...standard option for scientists by requiring graduate students to pay for part of their tuition. Having trainees...postdoc salaries be paid on fellowships instead of grants such that PIs no longer save grant money by paying... still be there without restructuring. Positive Points for Trainees - Taking Steps to Achieve a Successful... all traits you are likely to find in graduate students and postdocs. Trainees however, need to be made...

...that find events stressful. Video chatting makes it pretty easy to make 30 minute appointments with people...academia is run by grant funding. Learning to write grants quickly, efficiently and effectively is a valuable...valuable skill. There are many different kinds of grants that you can apply for. If you are in a lab, your...practicing and getting familiar with the process. Bonus points for being able to just look at a paragraph and ...Here are some sites that have compiled lists of grants: Science: Where to Search for Funding The Ohio...For Your Research University of Florida: Training Grants and Other Funding Opportunities L’oreal USA For...that you've been working on, let us know in the comments below! Additional resources on the Addgene blog...

...the pandemic, it’s hard to get these reagents. However, when reagents have been made available through a...because they provide rapid access to validated reagents, especially during a time when these materials...Everyone. Case study 1: Cell lines Many of the reagents used to investigate SARS-CoV-2 were developed ...endeavor. Case study 2: Plasmids Another example of reagents that have broad applications but have proven useful...plasmids and resources! Case study 3: Mouse strains Reagents developed during the SARS-CoV pandemic more than...from the scientific community. Deposits of new reagents, policies from journals and granting agencies ...for storing, validating, and distributing such reagents are all ways in which the scientific community...

... model. AAV plasmids contain several important elements, but among the most important are the inverted...inverted terminal repeats (ITRs). ITRs are the elements that make an AAV plasmid an AAV plasmid.  Within the... and initiate replication. ITRs are cis-acting elements, which means that they are in the same DNA molecule...types  Since ITRs are one of the most important elements of AAV and rAAV vectors, modifications can drastically...prevent unwanted mutations?  During propagation, mutants rapidly replicate in a growth culture and result... sequencing is also helpful in quantifying ITR mutants since analysis of individual reads allows for identification...the Addgene blog Viral Vectors 101: Viral Vector Elements Viral Vectors 101: Pseudotyping Viral Vectors ...

...muscle contractions) are produced by orchestrated movements of networks of proteins consuming molecular fuel...undergoing local conformational changes. These tiny movements are then amplified tens or hundreds of thousands...decades, and they were known to be resistant to detergents and degradation under a variety of conditions...see figure to the right), we identified R-body mutants that extend at both lower and higher pH. We initially...initially used this screen to find R body mutants that couldn’t extend at a normal pH and later realized...F. R. Organization and expression of genetic determinants for synthesis and assembly of type 51 R bodies...

...of the work, such as physically carrying out experiments like PCR and running gels, but in research you...in Ghana. My role involved teaching the nursing students and community health workers some basic biology...spreadsheets. It was great to be in the lab doing experiments  without the constant fear of failure because...company carries out clinical trials of drugs and treatments for external companies. It was really exciting... particular situation, communication between departments was sometimes difficult and the solution attempted... companies which are reliant on contracts with clients is that, when those contracts fall through or are...pancreatic cancer where one drug does not work for all patients, leading to very high mortality. I really loved...

...small errors that significantly impacts your experiments. What do you do when you’re in the lab and you're... steps like “check drawers for light-sensitive reagents” or “re-sterilize now.” Have a physical copy of...Have a physical copy on hand as you run your experiments. Try drawing out a protocol as you read it, or...an investment in accuracy.  Bench setup Small adjustments in the physical organization of your workspace...you track samples as you add them. Moving your reagents from your left side to your right as you use them...are endless.    Fig. 2: Blugene moves reagents from left to right as they use them.    Prevent...focus, which was extremely effective. Others ran experiments at odd hours, used more secluded workspaces, ...