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Showing: 801 - 840 of 1100 results

  1. Hot Plasmids and Viral Preps - March 2021

    Type
    Blog Post
    ...through our hot plasmids articles. These articles provide brief summaries of recent plasmid deposits and ... cells that make up the human body and G-baToN provides versibility in tracking and modulating the interactions...libraries encode target sites flanked by randomized nucleotides in place of a protospacer adjacent motif (PAM... viral aliquots from our plasmid collection to provide ready-to-use viral preps. Here are some of the ...

  2. Nanoblades: Tiny CRISPR Ninjas for Genome Editing Difficult Cells

    Type
    Blog Post
    ...respectively. Primary cell type Gene targeted by guide Editing efficiency hiPSCs  EMX1  67% Mouse BM...in cells electroporated with RNPs using the same guide. Nanoblades also had lower off-target effects when...insertion of a DNA sequence ranging from a single nucleotide change to large insertions like the addition ...Addgene.org Check out our comprehensive CRISPR guide Find CRISPR plasmids and resources Learn about viral...

  3. Fluorescent Proteins 101: Introduction to FRET

    Type
    Blog Post
    ...measure the tension in the helical structure of spider silk, or the activity of a protein in a synapse... observed light emission by fluorescence. The widespread use of FRET with fluorescent molecules, including...As the choice of fluorophore pairs is a key consideration for a FRET experiment, the possibilities can...PMCID: PMC3810607. 2. Bajar, Bryce T., et al. "A Guide to Fluorescent Protein FRET Pairs." Sensors 16.9...

  4. Experimenting in an Open Source lab: from CRISPR to Cats

    Type
    Blog Post
    ...bigger movement in a variety of research sectors to provide open source technologies and spaces where interested...One of the aspects of this project that makes it ideal for makerspaces is the data is freely available,...projects). This allows the project to be open to a wider group of people. This is a perfect starter project...equipment becomes obsolete amazingly fast, but with a wide range of skills among our members we’ve managed ...

  5. Degrading DNA with Cascade-Cas3

    Type
    Blog Post
    ...powerful than it is. The nuclease domain of Cas3 is considered weak, especially compared to the power of Cas9...Repetitive DNA reeling by the Cascade-Cas3 complex in nucleotide unwinding steps. Molecular Cell, 70(3), 385-394...Liao, M., & Ke, A. (2018). Structure basis for RNA-guided DNA degradation by Cascade and Cas3. Science, 361...CRISPR-Transposons Additional resources on Addgene.org CRISPR guide Cascade-Cas3 Plasmid collection ...

  6. Viral Vectors 101: Producing Your rAAV

    Type
    Blog Post
    ...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...

  7. 3D Printing Meets CRISPR Cas9

    Type
    Blog Post
    ...crystallographer working with Jane Richardson, provided an initial solution to this problem with the development...the surface of the protein where the nascent polypeptide emerges from the exit channel.   He then said...allow the complementary base pairing between the guide RNA and the target DNA strand, and then activate...User Resources on Addgene.org Visit Our CRISPR Guide Page CRISPR/Cas9 History Browse All CRISPR Plasmids...

  8. Tips for a 1st time AAV user (by a Rookie AAV user)

    Type
    Blog Post
    ...plasmids to produce AAV: 1) the RepCap plasmid which provides the AAV replication (rep) and capsid (cap) genes...primarily found in the culture media, you should consider performing PEG precipitation. Instead of sonicating... details. Pro tips Watch this AAV purification video! It didn’t exist when I did my first iodixanol gradient... gradient purification, but I wish it had. The video gives a brief overview of how iodixanol purifications...

  9. 10 Basic tips for mammalian cell culture

    Type
    Blog Post
    ...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...

  10. Interview: Nicola Patron on Plant Synthetic Biology, MoClo, and More

    Type
    Blog Post
    ...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...

  11. Drew Endy Introduces the Biobrick Public Agreement Plasmid Collection

    Type
    Blog Post
    ...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...

  12. Getting to Know Expansion Microscopy

    Type
    Blog Post
    ...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...

  13. Capturing a Moment with Marc Zimmer’s “The State of Science”

    Type
    Blog Post
    ...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...

  14. CRISPR Antimicrobials

    Type
    Blog Post
    ...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 ...

  15. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    ...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...

  16. Hot Plasmids Spring 2024

    Type
    Blog Post
    ... 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...

  17. Plasmids 101: Plasmid Incompatibility

    Type
    Blog Post
    ...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...

  18. CRISPR-mediated Plant Base Editors

    Type
    Blog Post
    ... 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...

  19. Viral Vectors 101: Pseudotyping

    Type
    Blog Post
    ...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 ...

  20. Rett Syndrome: A History of Research and Therapeutic Outlooks

    Type
    Blog Post
    ...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...

  21. Plasmids 101: E. coli Strains for Protein Expression

    Type
    Blog Post
    ...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...

  22. An Integrin Antibody Toolkit from IPI

    Type
    Blog Post
    ... 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...

  23. Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus

    Type
    Blog Post
    ...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...

  24. Plasmids 101: Common Lab E. coli Strains

    Type
    Blog Post
    ... 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...

  25. Recombinase-based State Machines Enable Order-dependent Logic in vivo

    Type
    Blog Post
    ...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...

  26. Antibodies 101 Series on TikTok

    Type
    Blog Post
    ...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...

  27. Overwhelmed? Take a Break With Our 5 Favorite Science Comics

    Type
    Blog Post
    ...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...

  28. Top Requested Plasmid of 2017 - pMD2.G

    Type
    Blog Post
    ...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...

  29. Addgene’s New Way to Help You Share Science

    Type
    Blog Post
    ... 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.  ...

  30. Deck the Lab 2022!

    Type
    Blog Post
    ...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...

  31. #DeckTheLab with Tinsel and Test Tubes, Fa La La La La La La La La

    Type
    Blog Post
    ... #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...

  32. Our New Antibody Mascot is....

    Type
    Blog Post
    ...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...

  33. Countdown to Halloween @Addgene

    Type
    Blog Post
    ...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.        ...

  34. Visualizing Your Plasmid Request Data

    Type
    Blog Post
    .... 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,...

  35. Newly Updated AAV Data Hub!

    Type
    Blog Post
    ...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...

  36. Plasmid Grant to Aid Cervical Cancer Screening in Ghana

    Type
    Blog Post
    ... 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...

  37. Podcast: Interview with Addgene's Founders

    Type
    Blog Post
    ... 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...

  38. Interview with Bart Newland, Addgene’s Newest Board Member

    Type
    Blog Post
    ...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...

  39. Addgene’s Blog Gets a New Look!

    Type
    Blog Post
    ...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...

  40. The Developmental Studies Hybridoma Bank: Over 25 Years of Antibody Sharing

    Type
    Blog Post
    ...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...
Showing: 801 - 840 of 1100 results