...subjected to quality control by Addgene before being distributed to customers. Details about our production protocols...control are described below. AAV Production AAV distributed by Addgene has been produced either in-house ...preparations are purified by iodixanol gradient ultracentrifugation and concentrated to purity and titers adequate...preparations is assayed by comparing the relative stoichiometric ratios of the viral capsid proteins VP1, VP2...containing lentivirus is first cleared by low-speed centrifugation and then by filtration through a 0.45 µm polyethersulfone...in 10% polyethylene glycol (PEG) followed by centrifugation. Precipitated pellets containing viral particles...

...commercial lateral flow strips, similar to pregnancy tests. With lateral flow strips, the presence of your...determined by the number of bands present on the strip. This type of readout allows for nucleic acid detection...detection almost anywhere as lateral flow strips are easy to transport and work rapidly, providing reliable...platform to detect SARS-CoV-2 using a lateral flow strip format. They published this in Nature Biotechnology... validation studies.     Jennifer Tsang contributed to updating this article. References Abudayyeh...

...Glove trick Putting on lab gloves can sometimes be annoying. To make it easier, try this quick trick. Just...Addgene has distributed over 800,000 plasmids since 2004. In that time, our lab has gotten a lot experience...Just flip, and press! Lab tip #2: Tube flick trick When combining reagents in the lab, it is important...several times. This will help ensure ensure equal distribution. Lab tip #3: Barcode everything Use barcodes...

...from 5,821 labs in 1,249 institutions across 55 countries.  Our first request was on September 8, 2004 (...pcDNA-p160MBP, if you're curious.) Since then, we've distributed 2,002,539 materials to 254,034 scientists in ...in 8,900 institutions across 111 countries. Just eight days into 2024, we've already shipped 2,078 items...scientists at 375 organizations in 21 different countries. It's safe to say we keep our shipping team busy...international community, we've partnered with distributors in China, Korea, Japan, India, and Brazil.  ...

...publication and we have already collected over 100 entries spanning six different species and dozens of different...tool hM4D(Gi) driven by a GFAP promoter in rat. Contribute your AAV data Just like Addgene’s plasmid repository.... We are grateful to the scientists who have contributed data so far and we encourage you to submit data...the data hub website as a thank you for your contribution to open science.  Please note that the AAV Data... currently only accepting data for materials distributed by Addgene.  Future directions As the AAV Data...

...This post was contributed by guest blogger Joachim Goedart, an assistant professor at the Section of ...several ways to measure stickiness. In vitro, ultracentrifugation or gel filtration is the most common analytical...mScarlet-tubulin (Plasmid #85045). We often use restriction enzymes in the multiple cloning site for generating...Bendezú et al., 2015) and the alpha subunit of heterotrimeric G-proteins (Janetopoulos et al., 2001, Adjobo-Hermans...Merel JW, et al. "Real-time visualization of heterotrimeric G protein Gq activation in living cells." BMC...Devreotes. "Receptor-mediated activation of heterotrimeric G-proteins in living cells." Science 291.5512...

...screen using PCR and a restriction enzyme digest.   Fig. 3: Restriction enzyme screening for resistivity...of your experiment for a screening strategy. Restriction enzyme screening is sensitive to single base ...changes, but the edit must occur at an existing restriction enzyme recognition sequence or generate a new... but your edit doesn’t affect any endogenous restriction enzyme sites? Never fear! You can introduce a...silent mutation should be designed solely for restriction enzyme sensitivity or resistivity, and thus be...

...This post was contributed by guest blogger Joachim Goedart, an assistant professor at the Section of ...genetically encoded fluorescent probe. Several factors contribute to the popularity of GFP including (i) fast and... mutation (A206K) is necessary to generate the strictly monomeric EGFP variant, mEGFP (Zacharias et al... bind the molecule of interest. For instance, pleckstrin homology domains can be used to detect specific...have successfully generated several functional heterotrimeric G-alpha subunit fusions that do not tolerate...Merel JW, et al. "Real-time visualization of heterotrimeric G protein Gq activation in living cells." BMC...

...forward to in 2023 and beyond. Last year, we distributed 165,000 plasmids, including 27,000 CRISPR plasmids...and general-use antibodies, is available in both trial- and full-size, with assay-supported recommended...our website saw 2.2 million visitors from 231 countries. Guides and protocols were our most popular content... million page views from visitors in over 200 countries. We, like everyone around the world, are learning...research community through sharing science and distributing high-quality materials and information. As a...

...When performing restriction digests on plasmids at Addgene, we sometimes observe something odd in our...various multimeric forms (e.g. concatemers, dimers, trimers, tetramers). Since then, multimerization has been...themselves cannot be used to detect multimers, since restriction sites in multimers usually repeat in tandem along...DNA without changing the size. As a result, a restriction digest of multimers will produce the same fragment...sample, you could try linearizing the plasmid via restriction digestion and then run the linearized plasmid...eBookAddgene's Molecular Biology ReferenceAddgene's Restriction Digest Protocol More resources on the Addgene...

...This post was contributed by guest bloggers and IMP organizers Rosario F. Godino, Rocío López-Diego, ...youth, our future, around the world.   In many countries of the world, professional internationalization.... Brain drain is a serious problem in several countries, however programs like IMP, bring closer than ...from talented professionals to their original countries to help build, together with the universities,...pre-program self-assesment. Achievements directly attributable to participation in the program included improvements... will also initiate mentoring pilots in other countries. In 2016, we expect the program to comprise over...

...nights, game nights, and trips to eat out together. “We also have a lab day trip each year and as we do ... the lab receives send-off via a gift, card, and trip to the pub. Dr. Ellis adds “we probably don’t celebrate...lot of yeast research, a few of those have been trips to breweries who happily exchange our expertise ...company outside of the lab by sharing meals, going on trips, and going to New Hampshire in the summer for the...

...through a distributor?  Yes! Packaged on Request service will be available through our distributor partners...of AAV plasmids will have technical or legal restrictions that make them ineligible for use in this service...international; varies widely by country and/or distributor.)  You should plan to receive your Packaged on...is launched, you’ll be able to work with the distributor to place a request and order if approved.  I ...

...on chemical (CaCl2, cationic liposome) or electrical (electric current forming membrane pores) transfection...virus. Virus purification can be a tricky process. You have to centrifuge the culture supernatant in order...The ability to infect non-dividing cells is not restricted to in vitro cell culture as lentivirus-derived...transducing non-adherent cells you might need to centrifuge your cells with the virus in order to aid viral...

...Restriction enzyme cloning is the workhorse of molecular cloning; however, one of its biggest limitations...that sequence modifications can only be made at restriction enzyme cut sites. The lambda red system is an... of DNA within E. coli and is independent of restriction sites. The lambda red system is derived from ...means cloning occurs in vivo, as compared to restriction enzyme cloning where the genetic changes occur...sections, key steps that differ from traditional restriction enzyme cloning will be explained in greater detail...screen for positive clones in most cases, and restriction enzyme digest can be used to screen plasmids ...

...This post was contributed by Kurt Swanson a structural biologist and protein engineer currently working...R206H point mutation on the receptor ACVR1. ACVR1 triggers appropriate bone regeneration in healthy patients...mouse models. As this biologic moves into clinical trials, there is new HOPE for AJ. But, as we in the scientific...fight against FOP in two ways. First, Addgene is distributing ACVR1 Plasmids from a variety of research labs...will be hosting the AJ’s Boston League team (Contributing many new heroes to the team in the process) ...

... a team to get a lot of stuff done.  Factors contributing to job satisfaction Being a good manager is ...mostly happy at work, they will work harder and contribute more. Assuming basic needs are being met (i.e...pay and benefits to live on), the factors that contribute to job satisfaction are a bit surprising. There...overwhelmingly agree that these are the factors that most contribute to job satisfaction: A feeling of being appreciated...organization and feel the importance of their contributions. Flexible work schedule Flexibility to manage...

...shut-down and the subsequent fluctuations in distribution. I don’t have space in this blog to call ...scientific community.  With the dramatic reduction in distribution, we had to figure out a way to maintain our ...operations until our normal work resumed. Addgene distributes and fulfills requests for scientists doing almost...especially proud to support Addgene now, when their distribution of plasmids for COVID-19 research is accelerating... cloning grade DNA service, and viral vector distribution is recovering from the pandemic-related slowdown...

...Human whole genome sgRNA iBAR library Article contributed by Susanna Bachle Figure 1: Initial steps...labeling using recombinase competition Article contributed by Aliyah Weinstein Sparse Predictive Activity...reporter for calcineurin activity in yeast Article contributed by Leah Schwiesow Calcineurin is a protein phosphatase...protein fusions as a DNA staining dye Article contributed by Jennifer Tsang Figure 4: Double stranded...single molecule-fluorescence assay to monitor restriction enzyme digestion of single molecules in real-...

... This post was contributed by guest blogger James D. Fessenden, an Assistant Professor at Brigham and... site when used in conjunction with Cy dye-nitrilotriacetic acid (NTA) conjugates (Kapanidis, Ebright,...50-69. Royal Society of Chemistry. 3. Ermolenko, Dmitri N., et al. "Observation of intersubunit movement...-mediated fluorescent labeling with (Ni2+: nitrilotriacetic acid) n-fluorochrome conjugates." Journal ...