...There are several facilities nationwide where you can obtain excellent quality, high titer rAAV (including...certain in vitro and in vivo applications, you can consider using unpurified AAV from the supernatant (Goodwin... in precision and variability, qPCR is the more widely used method due to the accessibility of instruments...101 series!  This post was originally written by Didem Goz Ayturk and updated by Ina Ersing in August 2023...

...of contamination for your experiments, the bright side is that you also have the power to take the added...working with so that cross-contamination can be identified. Even if some mammalian cells lines have similar...check under the light microscope will help you identify bacterial and yeast contaminants. Set up a routine...will help you avoid cross-contamination through accidental mixing and will also help you keep good records...

...wanted to expand the toolkit. We have included a wider range of terminators than are commonly used as we...rusts and we are collecting sequences that will provide a similar resource for those involved in monocot... really and we’ll be producing an instructional video soon to make it easier for people to use the Golden...think plant biology is massively underfunded considering how important it is. I would really like to reach...

...DNA systems capable of storing data and computing inside living cells. He's also a co-founder of the BioBricks...Champion of Change, a program created as part of President Barack Obama's Winning the Future Initiative. ...proteins. That's central to everything that goes on inside a cell. Until we get a handle on precise, forward...promise that we could instead have things like peptide or RNA aptamers that act as genetically encoded...

...Microscopy. What is Expansion Microscopy?  ExM provides a straightforward answer to the space problem....minimized through tweaks to these steps. But even under ideal polymerization and digestion conditions, loss of...Bournonville L, et al (2021) Expansion microscopy provides new insights into the cytoskeleton of malaria ...Sommer C, Rizzoli SO, Danzl JG (2019) A practical guide to optimization in X10 expansion microscopy. Nat...

...inclusion of some interesting case studies, and Zimmer provides several. In Part Three of the book, called simply... and her essential-but-unlauded role in the identification of DNA’s double helix. Perhaps to the dismay...dismay of middle-aged scientists, the identification of Green Fluorescent Protein (GFP) and its development...(for funding, etc.) with that produced by the confident and flamboyant Watsons.” In Part Four, “New Science...

...less potential to become resistant to? CRISPR may provide a method for doing just that. While challenges ...problem. Above all, we need new antimicrobials, ideally ones for which the development of resistance will... antimicrobials using efficiently-delivered RNA-guided nucleases.” Nature Biotechnol. 32(11): 1141-5. ...CRISPR Plasmids for Your Research Read Our CRISPR Guide Pages Find CRISPR gRNA Design Tools ...

...numbers, but the latter eliminates the step of identifying prey proteins after screening. It is also possible...activation. Including a prey-only control also provides a baseline level of reporter activation. Vary ... And most importantly: Independently validate identified binding partners with other techniques! Minimizing... While this may be difficult to resolve for a wide range of prey proteins, there are now a number of...

...pseudotyping allows these viruses to infect neurons. Besides altering the host tropism of your virus, there ... as a retrograde tracer so that scientists can identify the neurons that project to a given brain region... resources on the Addgene blog: Read the Quick Guide to All Things Lentivirus Get tips for titering your...Resources on Addgene.org: Check out the lentivirus guide Find ready-to-use lentiviral preps at Addgene ...

...that MECP2 binds to methylated CpG dinucleotides and CAC trinucleotides during neuronal maturation (Tillotson...Sinnamon et al., 2020). They introduced a mouse MECP2 guide RNA and a hyperactive version of the human RNA base...fine-tuning dosage so that enough wild-type MECP2 is made inside the cells, but this is exciting progress in tackling...

...theory for plasmid incompatibility is due to an identification issue: the CBPs in the cell can’t tell the ... lost from the cell at a higher rate than the coresident plasmid. Asymmetric plasmid loss occurs for several...based on sequence data. However, if these do not provide clarity, a wet lab transformation experiment will...Genomics of microbial plasmids: classification and identification based on replication and transfer systems and...

... defective gene, Xm, the sequence of which is identical to the correct gene, Xw, except for a single base...vectors have two Bsa1 sites to clone the single guide RNA (protospacer+scaffold) in a pol-III (OsU3) promoter... in a ‘GC’ sequence context. If this cannot be avoided, select the target AID version. Expect very low...PMC4873371.  2. Nishida, Keiji, et al. "Targeted nucleotide editing using hybrid prokaryotic and vertebrate...

... Adamson Lab recently used a CRISPRi screen to identify additional host factors impacting prime editing...subthreshold voltage fluctuations, while having high fidelity during action potentials of fast-spiking neurons... combination with miR124T targeting sequences, provides a simple yet valuable tool for microglia research...2023). Simple and Highly Specific Targeting of Resident Microglia with Adeno-Associated Virus. bioRxiv...

...growth phase are alive and healthy, which makes them ideal for protein expression. If you wait too long to ...and pLysE—the difference being that the latter provides tighter control of basal expression. What if I... growth media, or if your plasmid is low-copy, consider reducing the antibiotic concentration. Growth ...which contain hydrophobic regions or multiple disulfide bonds may aggregate and become insoluble. These...

...First, some history. In 2008, the Auricchio lab decided to solve the issue of “tiny vector, big transgene...of expressing a truncated protein. This can be avoided by excluding a stop codon from donor A in any of... to be reconstituted, there is currently no one ideal split vector system. Overall the efficiency of transgene...Resources on Addgene.org: Check out the Addgene AAV guide Find ready-to-use AAV preps from Addgene's viral...

... function — providing researchers new tools to identify integrin presence, study signaling mechanisms,...function-blocking integrin antibodies exist, almost all bind outside the ligand-binding pocket, their sheer size impeding...monoclonals, we anticipate that our chemically identical hybridoma-derived antibodies will also work in... More resources from Addgene Addgene's Antibody Guide Antibodies 101: Flow Cytometry Antibodies 101: Introduction...

... the thousands! The goal of this article is to provide enough background for you to distinguish the features...resistance to tetracycline Additionally, Table 2 provides a quick reference for some of the popular strains...strains are all based on E. coli K-12 and are considered BSL-1. Table 2: Lab strains of E. coli Strain...a Greek delta (Δ).  Additional resources We've provided an overview of the common lab strains; however...

...DNA-sequence configuration (state) based on the identity and order of a particular set of inputs, may be...sequencing.  A less straightforward, but more population-wide (as opposed to single-cell) approach is to interrogate...too expensive, then the qPCR method should be considered as an alternative. The qPCR method can be performed...researchers adapt GRSMs for their research, we provide MATLAB-based software that automates the design...

...updates whenever a new video is posted! If TikTok isn't your platform, all of our videos will also be shared...bite-sized portions of antibody concepts. Each short video answers an Antibodies 101 question, like "what is...

...different? If you still need an extra push, there’s evidence that laughter really can be good for you. A search...laughter” turns up a few fun and very recent bits of evidence. (What can I say, we are scientists). Researchers...way to connect with other people. (We don't need evidence for that one, do we?) OK, so, now that we’ve convinced...

...cells that it can infect. The VSV-G envelope is widely used and requested because it enables lentiviruses...how lentiviruses work, check out our lentivirus guide page. For more protocols and information, be sure...requested plasmid of 2018! Additional Resources Quick Guide to All Things Lentivirus 5 Tips for Troubleshooting...

... with “A Better Way to Share Science” for: The inside scoop from scientists who have deposited plasmids...technician, or Principal Investigator. How-to guides for troubleshooting your experiments and getting...and Facebook. We’ll track down the story, get the inside scoop, and keep you informed.  ...

...take samples from my assay at like 23:00, so I decided to make a giant snow angel for biologists in our...but to place it next to our lab trash (which coincidentally matches our Christmas tree's RGB colors) 😅...)   Here’s how to enter: Post your picture or video for either contest on Twitter or Instagram by December...

...After an agonizing week spent in cloistered consideration, and with much debate, we have finally narrowed...   But between these options, we simply can't decide! Abi, Lin, or Blottie? Which name best fits our...our new immunoglobulin friend? Please help us decide by voting for your favorite on this form or head over...

... #DeckTheLab contest is a time for scientists worldwide to show off their decorating skills. To join the...contest:   🎶Walking in a Winter Wonderlab🎶 We decided to #DeckTheLab with virus snowflakes from @CVRHutchinson...2020 Here’s how to enter: Post your picture or video for either contest on Twitter or Instagram by December...

...were indeed occurring earlier but that the photo evidence was destroyed to protect the innocent revelers...always nostalgia (old board games and movies can provide great character costumes). All of this costume ... checkout our FB page on Friday for more photo evidence.        ...

...My dad was a chemist and he actually gave me the idea of going into law. He said, “you know, the patent...my role is bringing experience and judgement to a wide variety of business decisions that are not “legal...see how I can help. Q: What do you enjoy doing outside of work? BN: I love to travel and that's been difficult...

...share scientific discoveries and developments, and provide tips and tricks for new and seasoned molecular ...specific area of interest. We also cover topics outside of these five areas and you can find what you need...comments below. We’ll track down the story, get the inside scoop, and keep you up-to-date. Be the first to...

.... Dashboards provide information in a concise and clear way that is accessible by a wide audience. The...parse through, aggregation of this information can provide valuable insight to the most requested plasmid,...

...too!   Using the Data Hub If you're trying to decide which AAV is right for your experiment, searching.... You can check out our step-by-step submission guide or email [email protected] to learn more. A great..., when published, will meet FAIR data-sharing guidelines. But what if my AAVs didn't work?  We love negative...

... founding Addgene, where the idea for Addgene came from and how the idea got off the ground Chapter 3 ...the Addgene Blog Addgene's a nonprofit? Get an Inside Look at Addgene's NGS Quality Control Addgene's...

... represents the continuation of our efforts to provide resources to researchers in developing countries...Ghanaian women by including PCR screening for HR HPV alongside the cytological tests currently in use. Gladys...screens. She hopes that these screens will both identify individuals who are infected with HR HPV and therefore...

...Studies Hybridoma Bank. Antibodies are the most widely used class of protein-binding reagents. In the ...and expanded. The clonal cells are screened to identify hybridoma lines, each producing a monoclonal antibody...ensuring affordable access to materials, repositories provide standardized quality-control and technical support...

...-one CRISPR/Cas9 vectors expressing up to seven guide RNAs (gRNAs). Sakuma said that the new system could...likely to work in animal embryos based on earlier evidence showing that a CRISPR/Cas9 plasmid could be injected...vector-mediated multiplex genome engineering. The system now provides an efficient targeting strategy for multiplex ...

...citation. If you’re considering requesting an Addgene antibody, the Data Hub can help you decide if an antibody... your application. It can be quite difficult to decide on which antibody to request for your experiment...

...student in the Collins' lab at the time) got the idea to engineer bacteriophage (viruses that infect bacteria...bacteria) for use as antibiotics. To develop this idea, Drs. Koeris and Lu founded the biotech startup ... as antibiotics 8:26 - 10:30 : How Dr. Koeris' decided to develop phage for use in a biotech startup 12...

...visited our labs in Cambridge, MA to learn about the wide variety of science careers that Addgenies represent...travels from 2015, including a great European photo slideshow from our UK team member Benoit Giquel. Bluegene...Regional High School Resources at Addgene  Read more Inside Addgene Blog Posts Get Career Advice from Our Bloggers...

...the DNA sequencing reaction should be repeated. Guidelines to help with DNA sequencing troubleshooting and... might suspect contamination in your sample or consider asking your sequencing facility to apply a special...vendors. The precipitation method has an unfortunate side effect of messing up the reaction around base 70...

...committee creates a compensation philosophy that helps guide employee compensation and also serves as an objective...think is the most important resource that Addgene provides the scientific community? Access to plasmids which...Addgene blog Learn more about about Addgene in our "Inside Addgene" blog series Subscribe to the Addgene blog...

...Stanford University). Right there on the floor outside the lecture hall. It was wonderful to see the ad-hoc...acoustic biomolecule-based dyes. This technology is widely applicable as bacterial expression of dyes allows...Microbial World’ showcased complex connections and ideas in various visualized approaches from hand-drawn...

...on the Addgene blog and have created and updated guides on various topics in biology over the past year...strategies for generating GFP fusion proteins. From considering protein size and shape, linker sequences, and...Cas9 and reverse transcriptase and a prime editing guide RNA, it can introduce insertions, deletions, and...

...Pooran surveyed a CRISPR google forum to get a better idea of how many people are having trouble, to determine...from Cornell University, is a bit old school and provides a thorough walkthrough of how you can use PCR ...Editing with CRISPR Cas9 Activators: A Practical Guide 10 Steps to a Perfect Science Talk Developing Lab...

...treatment, it is best to identify a cocktail of 3-12 phages. Once these phages are identified, they must then ... forward to help, the Phage Directory team must decide which labs should receive the clinical sample to...

...high-throughput manner, but these systems have not been widely employed in vivo. The CHIME and new X-CHIME systems...deposited at Addgene by Arlene Sharpe’s lab, allow for wide deployment of in vivo gene knockout in immune systems...CRISPR Cheat Sheat Resources from Addgene.org CRISPR Guide CRISPR: Protocol for Genome Deletions in Mammalian...

...may have only 2-3 formal supervisors that will provide guidance and experience. Navigating a fulfilling...about mentoring In this blog series, I plan to provide answers to the top questions scientists ask me ...teaches you to fish, doesn’t fish for you) Helps identify a Mentee’s strengths and make the most of them...

...Protein Collection webpage. These plasmids are divided into the following categories: Over 300 empty ...localizes in the cell, use these markers as your guide. As Michael Davidson told the Tallahassee Democrat...go together, we can fix it. Right now, we have no idea." Now, it will be that much easier for scientists...

...viewing sequencing traces, while the paid version provides more tools for plasmid mapping and design (Figure... deposited with Addgene. NCBI BLAST: Compares nucleotide or protein sequences to sequences in a database...molecular biology protocols Browse our protocol video collection ...

...know that the Fc domain provides structure, determines isotypes, and provides a place for secondary antibodies...yes, it does! While the Fc domain’s functions, besides structure and isotype determination, don’t generally...

...reporters that are detectable by an inexpensive and widely available imaging platform: ultrasound. Challenges...experiments because  ultrasound devices are inexpensive, widely available, and have deep tissue penetration and... imaging. Specifically, arg1 expressing E. coli aided in ultrasound imaging of the mouse gastrointestinal...

...Integrated DNA Technologies (IDT). “It was fairly accidental. I was finishing graduate school, and didn’t ...these days. I do some public speaking and a bit of video work for webinars (get tips for your next science...bring people in.” Are you interested in careers outside academia like science communication? If so, let...