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We narrowed to 1,025 results for: URE

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  1. What's New in CRISPR - March 2020

    Type
    Blog Post
    ...They generated these editors using the BE4max architecture. The new editors reported in this study have... levels relative to BE4max.  Read the paper in Nature Biotechnology Find the base editor plasmids CRISPRi...CRISPRi in Caulobacter crescentus Figure 1: Serial dilutions of CRISPRi in Caulobacter cells for...Enhanced CRISPR-based DNA demethylation Figure 2: The components of Casilio-ME1. Image from Taghbalout... methylation-silenced genes. Read the paper in Nature Communications Find Casilio-ME plasmids If you...

  2. Working 9-5: an Addgene Co-op

    Type
    Blog Post
    ...It was immediately clear that Addgene wanted to ensure applicants understood what the expectations were...responsible for helping other scientists on the team ensure the deposits we received are what the depositing...for me at Addgene. There is a clear effort to make sure people feel connected.    As I became accustomed...aided by the efforts being put in by Addgene to ensure their employees are feeling comfortable.     Lucie... Tips for Writing a Good Cover Letter 7 Tips to Secure a STEAM Internship This Summer  ...

  3. New Norepinephrine Indicators: nLightG and nLightR

    Type
    Blog Post
    ...nLightR (Kagiampaki et al., 2023).     Figure 1: Properties of nLightG and nLightR. A, Structural.... Scale bars, 10 μm (HEK293T), 20 μm (neurons). Figure and caption adapted from Kagiampaki et al., 2023...enhanced response time allows for more precise measurement of norepinephrine uptake throughout the brain...indicators for monitoring norepinephrine in vivo. Nature Methods, 20(9), Article 9. https://doi.org/10.1038...ColabFold: Making protein folding accessible to all. Nature Methods, 19(6), Article 6. https://doi.org/10.1038...

  4. A Better Way to Get Customer Support: The Help Center

    Type
    Blog Post
    ...called “self-service.” The idea here is to build features that allow users to help themselves, saving time... will continually update the Help Center to make sure that, whatever your support needs are, we’ve got...to the Help Center, you may have noticed a new feature on our website; towards the bottom right corner...www.addgene.org. We’re excited to make these new features available to the Addgene community, and we’d love...Read other Inside Addgene Posts Learn about New Features on Our Plasmid Pages Resources at Addgene Visit...

  5. Celebrating Outstanding CRISPR/Cas9 Achievements at the Dr. Paul Janssen Award Dinner

    Type
    Blog Post
    ...was delighted to be invited to the ceremony for surely what is one of many awards that will be bestowed...in markers on a whiteboard. The final image is pictured here…she started from a blank white slate and ...experimentation to the eventual translation to the clinic and cures. There is no doubt that companies are swarming to...computer to fight the known viruses. Bacteria have figured out how to download a bit of virus sequence to ...example, the technology is already available today to cure sickle cell anemia in stem cells but this can’t ...

  6. An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency

    Type
    Blog Post
    ...facilitating recovery of edited animals. Figure 1: In the background are 504 restriction digests... demonstrated that selection for repair of a temperature-sensitive pha-1 mutation significantly enriches...making screening for knock-ins much more efficient. Pure PCR repair templates are critical for efficient ...editing efficiency improved by using more dilute mixtures (Doksin et al., 2018). The Mello lab also reported...visit Addgene’s C. elegans CRISPR plasmids page. Future perspectives It is interesting to note that recipients...and PCR-based homologous recombination donors in cultured Drosophila cells. Nucleic Acids Research 42:e89...

  7. Fostering communication in remote collaborations

    Type
    Blog Post
    ...can be tricky, and it takes extra effort to make sure everyone in the meeting has an equal opportunity...emailing documents to one person at a time. Just be sure there is a point person responsible for incorporating...will come from the collaboration. Some things to figure out are timelines for experiments and manuscripts...from each other, access specialized equipment, or ensure that techniques are being performed the same way...feedback with everyone involved can speed up the future directions of the project. The state of research...

  8. Embracing Serendipity: A Crucial Element in the PhD Journey

    Type
    Blog Post
    ...discoveries, the world of academia often emphasizes structured methodologies and rigorous research plans. However...strategy could entail immersing yourself in diverse literature. By delving into new topics and expanding your...taking field trips, and spending time outdoors in nature or visiting museums can be excellent methods to... times (or use the notes app on your phone) to capture fleeting thoughts and connections that emerge during...recognizes marvel in all things.   Conclusion  In the structured world of lab research and PhD studies, where ...

  9. Early Career Researcher Toolbox: Free Online Molecular Biology Tools

    Type
    Blog Post
    ...molecular biology toolbox organized, today’s post features a list of free online molecular biology tools ...provides more tools for plasmid mapping and design (Figure 1). Benchling: While you might think of Benchling...Wayne Davis from the University of Utah.   Figure 1: A plasmid map created in SnapGene of Blue Flame...tools Addgene's analyze sequence tool: Annotates features, BLASTs, aligns, digests, and translates a sequence...access this tool, set up an IDT account. Figure 2: Addgene's Analyze Sequence tool.   DNA sequence...

  10. History of CRISPR Cas - A tale of survival and evolution

    Type
    Blog Post
    ...invading phages but to also destroy the phage upon re-exposure (2, 3). More specifically, specialized Cas proteins... immune system seems like the winning ticket to ensure prokaryotic survival; however, viruses, too, are...strongest will to survive. Genome engineering and future directions CRISPR/Cas inspired genome editing was..., Mojica FJM, Wolf YI, Yakunin JvdO, Koonin EV. Nature Reviews Microbiology. 2011 Jun; 9, 467-477. Bondy-Denomy...Bondy-Denomy J, Pawluk A, Maxwell KL, Davidson AR.  Nature. 2013 Jan; 493, 429–432. Cong L, Ran FA, Cox ...

  11. Fluorescent Biosensors for Measuring Autophagic Flux

    Type
    Blog Post
    ...because it turns a dynamic process into a static measurement and it provides limited information about what...fluorescent autophagy reporters now allows for the measurement of autophagic flux, or the changes in autophagic...strength excited at 586 nm to 440 nm serves as a measure of autophagy induction: when Keima is in the cytosol...is high. When untargeted, Keima can be used to measure bulk autophagy, but the addition of a mitochondria-targeting...18094608. Sargsyan, A., et al. "Rapid parallel measurements of macroautophagy and mitophagy in mammalian...

  12. Why Add Sucrose? Improved Yields for Adeno-associated Virus Preparation

    Type
    Blog Post
    ...2010). Rather than being released into the cell culture supernatant, the viral particles associate with.... Coroadinha, A. S., et al. "Effect of osmotic pressure on the production of retroviral vectors: Enhancement...overproduction of antibodies by applying osmotic pressure and sodium butyrate." Biotechnology and bioengineering...Shen, Chun Fang, and Amine Kamen. "Hyperosmotic pressure on HEK 293 cells during the growth phase, but ...serotype-dependent release of functional vector into the culture medium during adeno-associated virus manufacturing...

  13. Summer SciComm: Science Storytelling

    Type
    Blog Post
    ...introduction, and a resolution.   Fig. 2: The structure of a story - not just for fiction! Image credit...more. This could be by outright stating the big-picture problem, by asking your audience a simple question...’t need to be anything fancy; it just needs to capture your audience’s attention enough to make them decide... new questions are next or how this will impact future research or policy.  It is worth mentioning that...patients or other affected people, man/animal vs nature, anthropomorphized animals, or even seemingly unrelated...

  14. Antibody Neutralization Response Against Pseudoviruses Expressing SARS-CoV-2 Spike Protein Variants

    Type
    Blog Post
    ...we've been able to measure their relative ability to neutralize.”   Figure 2: Sera used in the...was not tested at the time of this study. Figure 1: Neutralization levels against spike pseudovirus...mutations was surprising,” says Balazs.   Figure 3: Neutralization outcomes of convalescent sera....V2 from neutralization by convalescent plasma. Nature 593:142–146. https://doi.org/10.1038/s41586-021...

  15. A New Optogenetic Tool Based on AraC Controls Gene Expression with Blue Light

    Type
    Blog Post
    ...🎶 “Bye bye L-arabinose drivePut your cultures in the shakerTurn the LEDs onAnd when you wantYou can ... a light-responsive domain that dimerizes upon exposure to blue light (Fig. 1). Another blue light-inducible...light-inducible dimerization domain, Vaucheria frigida aureochrome1 (VfAu1), led to functional BLADE variants, too...expression needed for most applications.   Figure 1: With blue light, the VVD domains of the BLADE...how spatially precise the activation is.   Figure 2: A bacteriograph of Michelangelo's "The Creation...

  16. Addgene Receives NIH BRAIN Initiative Grant to Create Open-Access Recombinant Antibody Resource

    Type
    Blog Post
    ... lines (hybridomas) that secrete antibodies in culture and provide renewable reagents with reliable and...plasmid-based and offer numerous advantages, including more secure archiving, unambiguous molecular definition, and...ready-to-use-proteins to facilitate use. Dr. Trimmer’s lab will ensure the quality of the antibodies using quantitative...Reproducibility: Standardize antibodies used in research. Nature 518:27–29 . https://doi.org/10.1038/518027a Bradbury... (2015) Antibodies: validate recombinants once. Nature 520:295–295 . https://doi.org/10.1038/520295b Bradbury...

  17. Another Pathway into Cells: iTOP

    Type
    Blog Post
    ...CPP-tagged protein and an untagged control could enter cultured cells and activate expression of a luciferase ...After iTOP treatment, both primary (hESCs) and cultured cells showed up to 70% resistance to diphtheria...enter the nucleus and cleave the specified target. Figure adapted from D’Astolfo et al. In certain instances...elsewhere in the genome. Second, expression is by nature transient, which may lower nuclease toxicity or...Techniques: Introduction of macromolecules into cultured mammalian cells by osmotic lysis of pinocytic ...

  18. Custom CRISPR Screens & the Green Listed Software

    Type
    Blog Post
    ...abilities, or other phenotypes that can be efficiently measured at the single cell level), are separated by, for...use ~400X more cells than gRNAs in a screen, to ensure that each gRNA has a good chance to contribute ...can be used: Design a screen with an easily measureable cellular phenotype (e.g. survival of a cancer...and minimize off-target effects of CRISPR-Cas9." Nature biotechnology 34.2 (2016): 184-191. PubMed PMID...and genome-wide libraries for CRISPR screening." Nature methods 11.8 (2014): 783-784. PubMed PMID: 25075903... parameters via a library-on-library approach." Nature methods 12.9 (2015): 823-826. PubMed PMID: 26167643...

  19. Addgene-Seeding Labs plasmid grant round 3: microbial drug resistance and biofilms, antidiabetic plant components, visualizing brain injury - and a glowing robot fish

    Type
    Blog Post
    ... susceptibility of biofilms formed on various structured surfaces. Plasmids expressing fluorescent proteins...deletions on biofilm formation and growth will then be measured to link the deleted genes to specific functions...several diseases. Her goal is to identify and isolate pure plant compounds with antidiabetic properties that...river, take water samples at specific locations, measure fluorescent signals from the sensor, and send a.... Subscribe to the Addgene blog for updates and future collaborations. Many thanks to all of the awardees...

  20. Crowdfight, a Platform to Boost Scientific Collaboration During COVID-19 and Beyond

    Type
    Blog Post
    ...research group in Scotland, who had just the right infrastructure and offered to collaborate with them. The partnership...cutting-edge research, is still one of the most valuable features of modern science in our view. And it was this...that prompted us to create Crowdfight.   Figure 1: Crowdfight is a community of scientists who ...collaboration and join efforts to shift scientific culture towards a more generous and rewarding way of making...receiving dozens of requests every day.    Figure 2: When a researcher needs a collaborator for a...

  21. Addgene Welcomes John Doench to the Advisory Board

    Type
    Blog Post
    ...through the literature, to go and talk to the people who actually have these plasmids, figure out what they...heard about RNAi at his MIT interview during a lecture given by Phil Sharp. After a few rotations, Doench... screening technology.” Doench thinks that the future of CRISPR will be in high throughput functional...resonates with me.” About Addgene’s role in the future, Doench says, Addgene is “a central resource for...

  22. Advice for Scientists Starting a Lab

    Type
    Blog Post
    ...postdoc come to an end and your next professional adventure begins. While many career pathways exist for scientists...ideas every day and decide on the best way to structure roles and responsibilities as we grew.” Initially...first hires will set the tone and atmosphere that future hires will look up to." She also recommends to ...rather than repel and hoard.” If you've already secured a faculty position, start studying up before you...process. Read Genevive Bjorn's "Ready, Set, Hire" on Nature Jobs for some quick advice and resources for hiring...

  23. Why and how to start studying chromatin (with Addgene's help)

    Type
    Blog Post
    ...it’s true that adding histones and chromatin architecture to every diagram portraying some aspect of eukaryotic...when I recently attended the 2018 Chromosome Architecture and Chromosome Organization Keystone Symposium...posters I saw at the 2018 Keystone Chromosome Architecture and Chromosome Organization. These aren’t the...noncoding RNA: NEAT1 RNA is essential for the structure of paraspeckles. Clemson CM, Hutchinson JN, Sara...posts below may steer you in the right direction! CAPTURE-ing Chromatin Interactions: Using CRISPR-dCas9 ...

  24. A Quick Guide to a Career in Software Product Management

    Type
    Blog Post
    ...information and form it into a roadmap--a plan for future features, programs, or tools to build. After organizing...my boss and coworkers, discuss current and future features being built for the analysis software program...simplest of terms, a software product manager helps ensure that the company or software group is building ...

  25. Cancer, Inflammation and Immunity - Harnessing the Body’s Defenses to Fight Cancer

    Type
    Blog Post
    ...inflammatory state by inducing immunosuppressive measures, for example the upregulation of the immunosuppressive... favorable state for tumor progression (Ref 2). Future treatments may need to target this immunosuppressive... analysis platform focused on analysing immune measures and correlating them with tumor mutations and ...locus with CRISPR/Cas9 enhances tumour rejection." Nature 543.7643 (2017): 113-117. PubMed PMID: 28225754...novel mouse glioma model using lentiviral vectors." Nature medicine 15.1 (2009): 110-116. PubMed PMID: 19122659...

  26. Multiplex Genome Editing with CRISPR-Cpf1

    Type
    Blog Post
    ...diagram below for an example of how this works. Make sure to order 5’ phosphorylated oligos or treat with ...plus a Cpf1 expression vector.  See the graph in figure 2 for a comparison of editing frequency resulting...plasmids are generally more efficient. Pro-Tip Make sure to reverse the orientation of the crRNA’s direct... used to multiplex edit 3-4 genes in a primary culture of mouse neurons and in vivo. For these experiments...Editing by CRISPR–Cpf1 Asing a Single crRNA Array." Nature Biotechnology 35.1 (2016): 31-34. Pubmed PMID: ...

  27. Addgene’s Viral Service - Why Virus? Why Now?

    Type
    Blog Post
    ...construction of our new BL2+ laboratory and the infrastructure capabilities to produce and distribute virus...laboratories all over the world. With this reach and exposure in the community, we saw the opportunity to leverage...distribution of viral samples. The Office Team made sure we could ship internationally and be ready to provide...formation of partnerships to expand the service and figure out what we can afford to do and when. Addgene’...the repository. The Outreach Team has been making sure scientists know about the new initiative and serving...

  28. Building and Validating Recombinant Antibodies: A Case Study

    Type
    Blog Post
    ...antibodies in adult rat brain. Figure from Andrews, et. al, Elife. Detailed figure legend here.     The ...chains as driven by two CMV promoters (orange). Figure from Andrews, et. al., Elife  A limitation ...the mAb becomes limited, which can be a threat to future reproducibility. The Trimmer Lab found that in ...recovering RNA from hybridomas grown in tissue culture, they were also able to isolate RNA from non-recoverable...

  29. Antibodies 101: Buffers, Storage, and Conjugates

    Type
    Blog Post
    ... The first thing you should do is record the manufacturer, lot number, and expiration date somewhere you...smaller than 10uL. You can leave them at room temperature when you’re working with them, but avoid freeze-thaw...okay, Blugene!    Buffers  You know what temperature to store your antibodies at, but it also matters... to react with other chemicals and biological structures, some of which you may need in your experiment...needed to excite them - and it’s a good idea to make sure that laser is available in your machine before you...

  30. GCE4All: Making Genetic Code Expansion Accessible

    Type
    Blog Post
    ... side chains, all of which can help determine structure and function relationships or, in more practical...non-canonical amino acid at the site of interest (figure 1). While the codon used is often one of the three...system (including media, cell type, and growth temperature) may need to be individually optimized for each...variety of ncAAs to study the relationship between structure and function…if you can get your protocol to work...focused on turning GCE tools developed in the literature into robust and universal techniques that researchers...

  31. How to Keep a Lab Notebook for Bioinformatic Analyses

    Type
    Blog Post
    ...printed for my physical lab notebook, along with any figures generated from the analyses: Goal: Before you ...will help you locate relevant information in the future. Approach: Outline a brief overview of the approach...repeating your work or making similar mistakes in the future. Stay organized Set up a table of contents in..., leave them in their own folder. Doing so will ensure that you are in fact using the same data for all...about the edit that was temporarily used. Just be sure that the original code remains in tact! Take notes...

  32. How to Make Friends and Meet People at a Scientific Conference

    Type
    Blog Post
    ...you meet will review your papers or grants in the future, and don’t be surprised to know that people think...about grants and projects that might help your work Tenure can depend on your national recognition and the...see enough of them).  Before the conference Make sure you have a business card – even students and postdocs... roll, say hello. Keep your cards handy and make sure your name tag is always visible. Some people learn...LinkedIn, especially if they are not in academia. Make sure to learn how to use LinkedIn properly. Then, follow...

  33. Visualizing Protein Turnover In Situ

    Type
    Blog Post
    ...I was studying a synaptic protein, I wanted to measure it’s levels in its normal neuronal context. In ...noncanonical amino acid tagging (FUNCAT, Dieterich et al. Nature Neuroscience, 2010) and allows the visualization...synthesized proteins in rat hippocampal neurons." Nature neuroscience 13.7 (2010): 897-905. PubMed PMID:...protein complexes in situ by proximity ligation." Nature methods 3.12 (2006): 995-1000. PubMed PMID: 17072308... of newly synthesized target proteins in situ." Nature methods 12.5 (2015): 411-414. PubMed PMID: 25775042...

  34. Supporting Rare Disease Research with the 2020 BeHEARD Awards

    Type
    Blog Post
    ...intellectual disability after Fragile X. There is no cure for CTD, which arises due to mutations in the SLC6A8...characterized by development delays, speech delays, seizures, and other symptoms as discussed in this educational...help the lab identify therapeutic targets in the future. Congratulations Mark Mellett! Siddharth Prakash... and is characterized by autism-like symptoms, seizures, and muscle weakness. So far, there have been ...patients with GNB1 mutations, the lab hopes to capture the signalosome of different receptors upon stimulation...

  35. Choosing the B(right)est Fluorescent Protein: Aggregation Tendency

    Type
    Blog Post
    ...There are a variety of methods that can be used to measure your fluorescent protein’s propensity to aggregate...likely in an antiparallel configuration), typical structures known as Organized Smooth Endoplasmic Reticulum...your protein of interest will be problem-free. In figure 1 we provide an example of the OSER assay with ... positive and negative controls for proper ER structure. Based on these data, we conclude that mKOk has...gathered can be shared by publishing it as (part of) figure 1, uploading it as a preprint, or documenting it...

  36. Summer SciComm: Preprints

    Type
    Blog Post
    ...or medRxiv.org) in the past few years.   Figure 1: Stand aside, CERN — it's a new age of accelerating... receive recognition for their efforts, inspire future research, and increase their impact. Open access... open scientific communication ecosystem Figure 2: The scientific ecosystem, enriched by open access...review is the only way to rigorously determine or ensure a manuscript’s quality. But even journal-based ...  Stern BM, O’Shea EK (2019) A proposal for the future of scientific publishing in the life sciences. ...

  37. Antibodies 101: Affinity Reagents

    Type
    Blog Post
    ...diabodies to be used for assembling protein nanostructures. Diabodies readily co-crystalize, a major advantage...only affinity reagent on this list derived from nature. Single-domain antibodies are extremely small at...but can have weak signals due to their monovalent nature. They are stable up to 80°C and can refold correctly...Books. Bates A, Power CA. David vs. Goliath: The Structure, Function, and Clinical Prospects of Antibody ...single-domain antibodies. Annu Rev Biochem. doi: 10.1146/annurev-biochem-063011-092449.             ...

  38. Transferable Skills Guide: Problem Solving

    Type
    Blog Post
    ...and designing future experiments. Being able to work through these types of bigger picture problems systematically...problem solving!  You might think that being able to figure out what went wrong with a PCR is a pretty niche...helping to resolve problems, but even so, I wasn’t sure how to best define this skillset. After sifting ...your problem solving skills can help convince a future employer that you are well equipped to work through...

  39. To Each HIS Own

    Type
    Blog Post
    ... to increase capture. Longer histidine tags are also recommended if you need a very pure sample. Because..., alternative fusion tags were used, such as S. aureus’ Protein A (Moks 1987). These tags, however, tended...denaturant. Not all proteins will refold once denatured, however, so take care when using this method....purification of human insulin-like growth factor I from culture medium of E.coli. Bio/Technology. 1987, 5:379-382...

  40. All in a Twist: dsRNA

    Type
    Blog Post
    ...mammalian cells. Here, we will cover how these structures are generated, why it’s important for cells to...proteins of interest. RNAi has even made it into agriculture as species-specific pesticides and insecticides...one strand of a dsDNA molecule; the 3-stranded structure produced is known as an R-loop. During transcription... transcription-coupled R-loops are transient in nature and will dissociate when gene transcription is ...vulnerable to various forms of DNA damage, and these structures can in turn lead to genome instability. Thus,...

  41. Career Coaching for Scientists: Why and Where Do I Find One?

    Type
    Blog Post
    ...career in science. Supervisors in the academic infrastructure are not always as effective in mentoring for.... Talk to the coach about their “style” and make sure this is a style that will help you. A good coach...also a very popular speaker and workshop leader. Lauren Celano of Propel Careers is not a professional ...afraid to ask and to make an investment in your future. Additional Resources on the Addgene Blog 6 Steps...Little about The Academic Labor Environment from the Future of Research Symposium Resources on the Addgene...

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

    Type
    Blog Post
    ...particles at the cell membrane. Mature Nanoblades then bud into the cell culture supernatant and can be purified...following four components into 293T cells. Figure 1: The key components of Nanoblades. Image from...iPSCs and BM cells after Nanoblade treatment as measured by their maintenance of pluripotency marker expression...Nanoblades loaded with Cas9-sgRNA ribonucleoproteins." Nature communications 10.1 (2019): 45. PubMed PMID: 30604748...

  43. Overcoming the Challenges of Lentiviral Production

    Type
    Blog Post
    ...expression of certain viral components; according the manufacturer these changes result in yields up to 25x higher...harvest is best for larger genes such as Cas9. See figure 1 for an example of how changing transfection ratios...While this option is attractive because the same procedure for a single 10 cm2 dish can be easily applied...consequently, this may not be the best method when pure virus is required downstream. When assessing lentiviral...titering your lentivirus Visit the viral vector featured topic page Resources on Addgene.org Lentiviral...

  44. Educational Planarian Tools for Teaching Developmental Biology Techniques

    Type
    Blog Post
    ...challenging for amateur scientists. Planarians, wormy creatures commonly found in freshwater ponds, provide a ...preparation of the tissue sample, several washes, and temperature changes to visualize the expression of specific... purified dsRNA with liver paste, and feed the mixture to the worms. The phenotypes of RNAi knockdown ...use these published protocols and want to share pictures, feedback etc. please write to [email protected]... double-stranded RNA in Caenorhabditis elegans. Nature, 391 (1998), pp. 806-811. PubMed PMID: 9486653....

  45. Hot Plasmids May 2018 - Optogenetics, Decaffeination, Biosensors, and Fluorescent Protein Tools

    Type
    Blog Post
    ... very little signal detected along the neurite (Figure 1.) Moreover, the group was able to demonstrate...are available at Addgene. Shemesh OA, et. al. Nature Neuroscience. 2017. PubMed PMID: 29184208.        ...methylxanthine drugs like theophylline. To facilitate future work with this goal, the Barrick lab has deposited...fLuctuation INcrease by Contact (FLINC)-based biosensors measure fluctuations of TagRFP-T fluorescence intensity...use to detect PKA activity can be generalized to measure phosphatase activity, second messenger activity...

  46. Lab Automation at Addgene

    Type
    Blog Post
    ...optimize more protocols in the future - something that will be key for our future growth. One of the biggest...pipette samples in and out of it, you can set up a procedure for it using the Biomek FxP. For example, we added...us to put a label on each tube by hand. We’ll be sure to tell you about more lab updates as we develop...have any great lab automation tips and tricks, be sure to share them in the comments section.   Additional...

  47. Split Fluorescent Proteins for Studying Protein-Protein Interactions

    Type
    Blog Post
    ...each part of a split fluorescent protein (FP) and measure the signal produced when the candidate proteins...interaction brings the pieces of the FP back together (Figure 1). This method is generally known as Bimolecular...Addgene is here to help!  General Design Figure 1: Two proteins are fused to the FP(11) and FP(...interact, the FP fragments can assemble the full structure of the fluorescent protein, producing fluorescent...number of strands in the canonical beta-barrel structure. Most frequently, the proteins are split between...

  48. Simple CRISPR-based Epigenetic Editing: dCas9-directed DNA Demethylation

    Type
    Blog Post
    ...modifications). Ready to get started?   Figure 1: Break free from the ball and chain!   Epigenetics... lot of things were cool: Walkmans, Back to the Future, and big hair. To what was perhaps a slightly smaller... (Hrit et al. 2018, Kaas et al. 2013). Figure 2: CRISPR TET-based epigenetic editing. In reality...expression (Sapozhnikov and Szyf 2021).    Figure 3: A simpler approach - dCas9 blocks DNMT1 methylation...some human cancers from their normal counterparts. Nature 301, 89–92. https://doi.org/10.1038/301089a0. Galonska...

  49. Adapting Toehold Switches to Detect Zika Virus

    Type
    Blog Post
    ...format that was stable for up to a year at room temperature (Pardee et al, 2014). As a demonstration of the...switches and amplification primers. This helped make sure that we had a number of successful sensors to draw...switch-based RNA sensors within an extensive infrastructure of molecular and electronic companion technologies...without purification or concentration of the virus. Future work Going forward, our short-term efforts are ...rapid prototyping and sensor assembly pipeline for future emerging pathogens or health crises. Many thanks...

  50. 5 Tips for Troubleshooting Viral Transductions

    Type
    Blog Post
    ...DH5α, grown at 30 °C instead of 37 °C. If you’re unsure about the integrity of your virus, perform a diagnostic...ultracentrifugation, you can reduce the volume of cell culture medium on the packaging cells immediately following...5 mL medium on a 10 cm plate). By reducing the culture volume into which the virus is released, a more...experiments later. Stay tuned to the Addgene blog and be sure to check out our viral vector guide pages to get...DNA and hepatitis C virus RNA quantification as measured with branched-DNA technology." Journal of clinical...
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