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We narrowed to 929 results for: KIN

Showing: 451 - 500 of 929 results

  1. Plasmids 101: Golden Gate Cloning

    Type
    Blog Post
    ...the design of flanking overhangs. Although there are theoretically 256 distinct flanking sequences, sequences...their recognition sequence, creating four base flanking overhangs. Since these overhangs are not part ...clone up to 7 gRNAs into one destination vector, making multiplexing easy.  Disadvantages of Golden Gate...

  2. Antibodies 101: Stripping and Reprobing Western Blots

    Type
    Blog Post
    ...by starting with the least-abundant protein and working toward the most abundant. If the abundance of your...precipitates that permanently stain the membrane, making that section of the blot unusable for reprobing...ll want to check for the primary antibody by re-blocking your membrane and re-incubating with your secondary...is the one time in western blotting that you’re looking for a “blank” result! If some signal remains, you...

  3. Plasmids 101: Modular Cloning

    Type
    Blog Post
    ...another optimized platform. Gene Stacking As the name implies, gene stacking is the use of a combination of...engineering complex cellular behaviors requires making complex manipulations to cells. For example: in...reagents and strategies are not available to those working with yeast or plants. Using MoClo, or better yet...

  4. Corralling and Cornering the Goldilocks Antibody

    Type
    Blog Post
    ... one place... well, lucky for you one is in the making by Dr. Zhuhao Wu and his colleagues! Dr. Wu and...for low-cost, high-quality antibodies for IHC. Working with collaborators and support from the Brain Research...enable scientists to have an intuitive guide for checking their staining and neuroanatomy without the fear...by submitting data, sending antibodies, and/or working with Dr. Wu please contact us here! Ava Shipman...

  5. Split Fluorescent Proteins for Studying Protein-Protein Interactions

    Type
    Blog Post
    ...localization in cells, or even single-particle tracking of the bound complex. There are so many FPs available...TagBiFC technique allows long-term single-molecule tracking of protein-protein interactions in living cells... can even be bright enough for single-molecule tracking in live cells.  Controls  When designing experiments...editing (Feng Zhang Lab 2015)  Reference Feng, S., Sekine, S., Pessino, V. et al. Improved split fluorescent...

  6. Plasmid-based Recombinant Monoclonal Antibodies: What They Are and Why You Should Be Excited About Them

    Type
    Blog Post
    ...  Second, plasmids are easy to store and share, making recombinant antibodies the most practical choice... and producing antibodies from plasmids is much kinder to our animal friends! Finally, if we want better...improve genetically-encoded research tools, it is shocking that the plasmids and sequences for antibodies...10.1038/520295b Bradbury ARM, Trinklein ND, Thie H, Wilkinson IC, Tandon AK, Anderson S, Bladen CL, Jones B,...

  7. Hot Plasmids and Viral Preps - March 2021

    Type
    Blog Post
    ...Epigenetic engineering with single-pot assembly Tracking cell-cell interactions New auxin inducible degron...Epigenetic engineering with single-pot assembly By Eric Perkins Though it’s easy enough to contemplate DNA in terms... own engineering plasmid tools in about a week. Taking advantage of Golden Gate cloning, the Haynes lab...human body and G-baToN provides versibility in tracking and modulating the interactions between cells....

  8. Early Career Researcher Toolbox: Finding Relevant Papers

    Type
    Blog Post
    ...tweet in order to easily access it later. Pro tip Checking the number of retweets, likes, and comments on...recent papers that the scientific community is talking about and jumping into the discussion! Semantic...altmetric score, is also displayed in your feed. By clicking on a paper in your feed you can see suggested ...Career Research Toolbox blog posts on free tools for making scientific graphics post and social media for scientists...

  9. Adeno Associated Virus (AAV) for Cell and Gene Therapy

    Type
    Blog Post
    ...This disease is caused by mutations in the common cytokine receptor gamma chain (c) and is characterized ...deficiency, Cystic fibrosis, Muscular Dystrophy, Parkinson’s disease, and HIV. A notable landmark study showed...AAV-based gene therapies are promising, scientists are working to overcome key challenges to their efficacy:  ...AAV vectors used to treat them. Scientists are working to overcome this issue by developing new capsids...

  10. Multiplexed Capture of Promoter-enhancer 3D Chromatin Structures Using CRISPR

    Type
    Blog Post
    ...Hi-C (Han et al., 2018). Think of 3C methods as taking a snapshot of chromatin looping with the photo ...CAPTURE method is similar to other 3C methods: crosslinking captures chromatin interactions, a restriction...together to create loops of DNA. This is key to linking pieces of DNA that are far away from each other...Zhang, Y., Liu, Y., Liu, N., Botten, G.A., Cao, H., Orkin, S.H., Zhang, M.Q., & Xu, J. (2020). Multiplexed...

  11. Fluorescent Proteins 101: Fluorescent Protein Timers

    Type
    Blog Post
    ...Terskikh et al.) and even displays predictable kinetics over 14 hours of expression in C. elegans embryos...Miyatsuka et al.) Distinguish between models for trafficking to the lysosome by fusing Medium-FT to LAMP-2A...structure and a particular biological event (trafficking to a subcellular location, start of gene expression...change color from blue to red report on cellular trafficking." Nature chemical biology 5.2 (2009): 118-126...

  12. Truncated gRNAs for Regulating Gene Expression

    Type
    Blog Post
    ...the development of a simple and scalable means of making targeted genetic edits. Cas9 is a DNA binding protein...Traditional Cas9 regulators In addition to its utility in making targeted modifications to DNA, Cas9 can be reprogrammed...potent Cas9-based transcriptional regulator without making any functional changes to Cas9. For instance, when...PMC4747789. 6. Platt, Randall J., et al. "CRISPR-Cas9 knockin mice for genome editing and cancer modeling." Cell...

  13. Production of Virus in Insect Versus Mammalian Cells

    Type
    Blog Post
    ...Whether you’re new to in-house virus production or looking to improve your current methods, understanding ... add an additional viral passage. This involves taking your first round of virus, called P1 (or sometimes... and may need to be optimized for your purpose. Making your decision The final decision of which system...10430340260355347. Vaughn, J. L., Goodwin, R. H., Tompkins, G. J., & McCawley, P. (1977). The establishment...

  14. Twenty Years of Addgene Sharing: CRISPR

    Type
    Blog Post
    ...editing!  CRISPR functions in use But exactly what kind of editing are people doing with all these CRISPR... modifier. We used these categories to see what kinds of CRISPR tools people were depositing and requesting... share useful tools, and as such, we’re always looking to understand our community better. We therefore...year, or approximately 104 plasmids shipped per working day, including appropriate MTAs and customs forms...

  15. Chromoproteins: Colorful Proteins For Molecular Biology Experiments

    Type
    Blog Post
    ...The ocean is full of striking visuals from fluorescent jellyfish, bioluminescent phytoplankton, and colorful... see if your cloning experiment worked just by looking at the cells on the plate. For example, if your... identification is a good place to start before taking on more time consuming and expensive methods. While...Learn more about FRET Find protocols and tips for making plasmid constructs Resources on Addgene.org Find...

  16. Academic vs. Industry Postdocs

    Type
    Blog Post
    ...enter industry since I was drawn to the idea of working as a team to develop therapies that could directly... since you can’t afford to spend too much time working on something extremely risky or exploratory. Getting...it does not divulge any propriety information. Working as a team: Industry is all about teamwork. A single...environment, surrounded by lab members who are all working on related topics, scientific discussion is often...

  17. Plasmids 101: Optimizing Plasmid Yields

    Type
    Blog Post
    ... unknown, it is entirely possible that you are working with a low copy plasmid. When increasing the culture..., or a home-brew optimized for plasmid yields. Spiking your media with supplements such as magnesium salts...can optimize your growth time, temperature, and shaking speed to maximize cell density and plasmid yield...especially those in deep-well plates, require a faster shaking speed for proper aeration (260-300 RPM). 37℃ doesn...

  18. Transgenic Organisms, Cas9 Gene Drives, and Appropriate Safeguards

    Type
    Blog Post
    ...Institute and Harvard Medical School. Scientists making transgenic organisms with Cas9 should be aware ...should be conducted with extreme caution. Why does making transgenic organisms with Cas9 risk creating a ...and single guide RNAs (sgRNAs) with appropriate flanking homology into a germline cell can create an RNA-guided...mosquito species. Reproductive confinement involves working with laboratory organisms that can't reproduce ...

  19. Plasmids 101: Multicistronic Vectors

    Type
    Blog Post
    ...accurate, as these peptides are thought to function by making the ribosome skip the synthesis of a peptide bond...cleavage efficiency. How do I get started? If you are looking to co-express your gene of interest along with ...with a plasmid that has multiple cloning sites flanking the multicistronic element(s), or you could replace...within the same multicistronic vector, effectively making a construct that expresses multiple unique genes...

  20. AAVs in Retinal Gene Therapy

    Type
    Blog Post
    ...11326284. 4. Bennett J, 2017. Taking Stock of Retinal Gene Therapy: Looking Back and Moving Forward. Molecular...pioneer in the field, Dr. Jean Bennett started working on ways to deliver genes to correct genetic conditions.... Jean Bennett (UPenn) that resulted in the groundbreaking 2001 study showing that AAV-mediated delivery...

  21. Getting to Know Expansion Microscopy

    Type
    Blog Post
    ..., are right around 250 nm, making it difficult to study the inner workings of the synapse by conventional...caused by loss of protein during digestion, or masking of epitopes during sample anchoring and gel polymerization...et al. 2019). Perhaps it’s the tool you’ve been looking for too!     References and resources References...

  22. Analyzing CRISPR Editing Results with ICE from Synthego

    Type
    Blog Post
    ...each sample by clicking on the sample name or on its corresponding bar graph entry. Clicking to initiate ...engineering has revolutionized the gene editing field by making experimental workflows considerably easier, faster... download the entire analysis as a ZIP file by clicking the Download Analysis Data button on the bottom...

  23. Sonic Hedgehog? Sleeping Beauty? Learn About the Genes Behind Addgene’s Conference Room Names

    Type
    Blog Post
    ...such as cell movement/motility, development, cytokinesis, and muscle contractions (think skeletal muscle...contexts within the fly including oogenesis and cytokinesis. Why name this protein after a vegetable substitute...encoded by spaghetti-squash, a gene required for cytokinesis in Drosophila." Cell 65.7 (1991): 1177-1189. ...resources on the Addgene blog Read our Quick Guide to Working with Drosophila Read our Plasmids 101 blog series...

  24. Plasmids 101: Plasmid Incompatibility

    Type
    Blog Post
    ...replication by binding to the origin of replication and blocking replication machinery (del Solar, 1998). As the... on each plasmid and pair the plasmids together (akin to sister chromatids coming together in eukaryotic...centromere-like region, and the NTPases end up “walking” the same plasmids to one daughter cell, instead...occurs for several reasons, including one plasmid blocking replication of another and one plasmid outcompeting...

  25. Bioinformatics at Addgene

    Type
    Blog Post
    ...process was working as expected, we moved forward with creating the next process and linking the output...needs of the user. Thus, a key criteria we were looking for was the ability to re-use or borrow existing... end of the script or by opening the files. By working on one process at a time, and testing a small set...

  26. New Acoustic Reporter Genes: Ultrasound Imaging of Gene Expression

    Type
    Blog Post
    ...as well as high spatial and temporal resolution, making it ideal for visualizing gene expression in vivo...ultrasound contrast. We (the authors) are currently working on improving and optimizing ARGs, with Marjorie...They also did not scatter ultrasound nonlinearly, making them difficult to distinguish from background tissue...intraperitoneal doxycycline injections to enable their tracking with ultrasound imaging (Fig. 2d). Creating a ...

  27. Antibodies 101: Reading a Flow Plot

    Type
    Blog Post
    ...stem cell research initiative at the King Abdulaziz University, Kingdom of Saudi Arabia. Bioinformation, ...markers are read in parallel.  Let’s imagine we’re looking for cells in a sample that are negative for marker...controls. You can usually tell where this point is by looking at the different populations. In the case of Figure...

  28. 5 Reasons to Use Reddit for Science Communication

    Type
    Blog Post
    ...appealing, but also scary. I do a lot of public speaking where I enjoy communicating face to face with ...early and brush up on best practices for LinkedIn networking. I was an early adopter of the LinkedIn blog ...ve said this before, but I also highly recommend taking every opportunity you get to speak publically about...Share Your Science 6 Tools & Tips: Online Social Networking for Scientists Sharing Your Lab Protocols: Using...

  29. pCXLE toolkit: Efficient episomal plasmid-based method to reprogram peripheral blood cells to iPSCs

    Type
    Blog Post
    ...collecting fibroblasts, which require a more invasive skin biopsy procedure. This becomes a major benefit when...patients, specifically children and patients with skin disorders for research and clinical trials, as it...usually require about 5-10 passages. Furthermore, working with Sendai virus requires stringent biosafety ...Road Ahead." Journal of Alzheimer's disease & Parkinsonism 6.5 (2016). PubMed PMID: 27882265. PubMed Central...

  30. Replacing Paper: Tips for Choosing an Electronic lab Notebook

    Type
    Blog Post
    ...situation differently, some adopting simple note-taking tools, others using excel sheets or organizing ... on the market today, ranging from general note taking tools, to more scientific field-based solutions...of use of the software. We identified that when talking about ELN costs and limited budgets, “a large percentage...ELN will support that. In addition, if you are working for a large institution or company, site licenses...

  31. Five Popular Model Organisms, Part 2

    Type
    Blog Post
    ...Iannaccone et al., 2009). Rats are larger than mice making them more useful for certain studies that involve...genetics and developmental patterns are conserved, making them great models for human disease research. One...eggs are large, opaque, and produced in abundance, making them useful for in vivo studies. These attributes...flowers. Image from BlueRidgeKitties. If you are working with Arabidopsis check out the GreenGate Toolkit...

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

    Type
    Blog Post
    ...fit a big gene into a tiny vector like AAV? By breaking the transgene into smaller pieces. Reassembly ...following transduction.  So what’s going on here? The working model is that the oversized AAV genomes are fragmented...Surveillance Mechanism That Eliminates Transcripts Lacking Termination Codons. Science 295:2258–2261 . https... Hirsch ML, Li C, Bellon I, Yin C, Chavala S, Pryadkina M, Richard I, Samulski RJ (2013) Oversized AAV...

  33. Antibodies 101: Introduction to Immunofluorescence

    Type
    Blog Post
    ...experiment. What is immunofluorescence? Generally speaking, IF is an application in which antibodies that... different approaches to be familiar with when talking about IF, direct and indirect. For direct IF, researchers...different colors. Multiplex immunofluorescence Speaking of different colors, having a palette of secondary...permeabilization, antigen retrieval, and sectioning. Blocking Block is used to reduce non-specific binding...

  34. Viral Vectors 101: Integrase-Deficient Lentiviral Vectors

    Type
    Blog Post
    ...forever!  This can be an advantage for researchers looking to generate long-lasting changes in cells. However...inserting viral DNA into the host genome. Without a working integrase, IDLVs can still deliver their genetic...Considerations for IDLV use If you’re reading this and thinking “Gee, I’d love to give these a try myself,” look...Linch, D. C., Bayford, J., Brown, L., Quaye, M., Kinnon, C., Ancliff, P., Webb, D. K., Schmidt, M., von...

  35. Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

    Type
    Blog Post
    ...Brainbow labeling has also been applied to cell marking and lineage tracing in other systems, such as zebrafish...directly trafficked to cell membranes. Membrane trafficking of farnesylated derivatives enables labeling ...fluorescence due to the addition of translation-blocking STOP cassette immediately following the promoter...dev.094631. PubMed. Multiplex cell and lineage tracking with combinatorial labels. Loulier K, Barry R,...

  36. Viral Vectors 101: Systemic Capsids

    Type
    Blog Post
    ...this post, we didn’t come across a broadly used working definition of systemic capsids. After some consideration...Beadle, K., Jang, M.J., Kim, H.M., Rajendran, P.S., Tompkins, J.D., Shivkumar, K., Deverman, B.E., Gradinaru...Chen, X., Wolfe, D. A., Bindu, D. S., Zhang, M., Taskin, N., Goertsen, D., Shay, T. F., Sullivan, E. E.... I. G., Pacouret, S., Chen, A. T., Chan, Y. A., Ryvkin, D., Gu, C., & Deverman, B. E. (2022). A high-efficiency...C., Lagerborg, K. A., Tellez, L., Krunnfusz, A., King, E. M., Ye, S., Solomon, I. H., Tabebordbar, M.,... Tabebordbar, M., Lagerborg, K. A., Stanton, A., King, E. M., Ye, S., Tellez, L., Krunnfusz, A., Tavakoli...

  37. CRISPR 101: Off-Target Effects

    Type
    Blog Post
    ...scenarios below for a CRISPR experiment which involves making specific knock-out edits in a gene(s) of interest...off-target frequency of 5%. Your experiment involves making a large sgRNA library to thousands of genes and...certain Cas enzymes are less compatible with the bulkiness that accompanies mispairing of bases. To reduce...enzyme swaps will only reduce off targets where nicking/cleaving of DNA occurs, not for deadCas activator...

  38. Antibodies 101: Multiplex Immunofluorescence

    Type
    Blog Post
    ...counting the number of puncta in each cell. When making figures for a paper or presentation, make sure ...binding non-specifically – try optimizing your blocking and washing conditions or consider using a different...concentrations of all the different antibodies, blocking, washing, and sample preparation steps, in order...Immunofluorescence Staining. In W. H. Yong (Ed.), Biobanking (Vol. 1897, pp. 299–311). Springer New York. ...

  39. Viral Vectors 101: Optogenetic Tools

    Type
    Blog Post
    ... channel with fast kinetics may be required, whereas a channel with slower kinetics can be used for lower...each light source used.   Channel and activation kinetics Some channels turn on and off very rapidly in ...thorough characterization data, like spectral and kinetic activation information, in the supplemental data...

  40. Deep Dive: Statistical Tests (Comparisons)

    Type
    Blog Post
    ... single dependent variable. I know what you’re thinking – this really sounds like a two-way ANOVA. And...calculated p values from smallest to largest and ranking them (1 being the smallest value). It calculates... you want to know type of distribution you are working with, graphing data with a histogram is sometimes... friendly neighborhood biostatistician happy by making your statistical analysis plan when you design ...

  41. Introducing the Addgene Antibodies 101 eBook!

    Type
    Blog Post
    ...eBook collection is a handy resource for anyone looking to use antibody-based applications in their research...: What is an Antibody” quite useful. If you’re looking to design your first flow cytometry experiment,...the Antibody 101 eBook is a great place to start looking for answers.    We hope you find the latest addition...

  42. Special Delivery: Fluorophore Targeting for FRET Studies

    Type
    Blog Post
    ...Miyawaki, Griesbeck, Heim, & Tsien, 1999), monitoring kinase activity (Ni, Titov, & Zhang, 2006), quantifying...pressure-test your FRET assay to make sure it is working? This article addresses these questions, which ...s best to consider these cases separately when talking about the tools that are available. Let’s consider...Denis V. Titov, and Jin Zhang. "Analyzing protein kinase dynamics in living cells with FRET reporters." ...

  43. New FLIS Plasmid (April Fools 2022)

    Type
    Blog Post
    ...Fly-related Issues in the Lab A significant drawback of working with fruit flies is their ability to escape their...experienced the minor yet consistent annoyance of walking into a fly lab and finding ourselves accosted by...system in a video filmed at Addgene:     If you're working in fly research, I hope you find this new technology...

  44. Addgene at SfN23!

    Type
    Blog Post
    ...viral vector specialist, Rebecca. They will be working with Andrew, the head of our scientific support...Specifically, Jo, Brook, and Angela are co-leading "Making a Greater Impact Using Clear and Accessible Scientific..., Instagram, and Bluesky.   Finally, if you're looking to brush up on your conference skills, check out...

  45. A Needle in a Base-Stack: Cas9 Structural Biology

    Type
    Blog Post
    ...   Unbound Cas9 Now that we know what we’re looking at, let’s peek at unbound—“apo”—Cas9 from Streptococcus... the absence of guide RNA, preventing Cas9 from making off-target cuts.   Pro tip! X-ray crystallography...region between the two (Jiang & Doudna, 2017). By linking the two nuclease domains, Cas9 ensures both are...biology can help you visualize the system you’re working with and better understand how to manipulate it...

  46. CRISPR 101: Cytosine Transversion Editors

    Type
    Blog Post
    ...chemically modify target bases without breaking the DNA backbone, making them efficient and precise tools for... So how have scientists solved this problem? By taking advantage of repair pathways within the cell — ...mammalian cells Researchers in the Xueli Zhang lab were making similar observations around the same time. (The...

  47. 5 Science Rap Videos & Their Creator: Tom McFadden

    Type
    Blog Post
    ...If you’re new to plasmids, we highly recommend checking out the video). Tom has made many more Science...sciencewithtom.com. Many thanks to Tom McFadden for taking the time to sit down and talk with us. Tom McFadden...McFadden is a YouTuber and science educator looking for creative ways to implement the next generation science...

  49. Summer Fun at Addgene!

    Type
    Blog Post
    ...common techniques and concepts. But what if you’re looking for information beyond the 101? That’s where our...our YouTube channel and watch it now! (Bonus: Tim kindly offered to finishing answering all submitted questions...check out our Instagram all summer long to see what kind of shenanigans we get into!  We hope you are as ...

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

    Type
    Blog Post
    ... used to thinking about the downstream results of actions. I’m also pretty adept at thinking about risks...was in the early 80s, and biotech was really just taking off. I found out that patent attorneys with a biology...
Showing: 451 - 500 of 929 results