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Showing: 421 - 450 of 954 results
  1. Minigenomes - a Safe Way to Study Dangerous Viruses Like the Ebola Virus

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    Blog Post
    ...is a nonsegmented, negative sense RNA strand approximately 19 kb in length. It contains short promoter ...replicated and transcribed in the cell when the appropriate viral proteins are provided in trans (see Other..., there are a few reasons arguing against this approach. First, this would prevent the study of the proteins...activity (4). To ensure that you end up with appropriate plasmid ratios, we suggest: 500 ng NP, 500 ng...
  2. Your Lentiviral Plasmid FAQs Answered

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    Blog Post
    ...lentiviral plasmids created for a variety of applications including cDNA expression, shRNA-mediated knockdown...only infect dividing cells. When choosing the appropriate gene delivery system, the specific target cell...type or tissues; this is useful in gene therapy applications where a distinct subset of the cellular population...free to post them in the comments and we would be happy to answer them! Note: A. Max Juchheim, Marcy Patrick...
  3. Technical Design of a Western Blot

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    Blog Post
    ...it includes validation data relevant to your application — and double-check that lot number while you’...unstained set of standards that covers a size range appropriate for the target(s) and transfer method. Membranes..., make sure to choose a chemilumescence kit appropriately sensitive for the amount of protein you are ...reference table to help you quickly decide what approaches best suit your experimental needs. Table 3: Optimizing...Increased time; increased reagents Good luck and happy blotting! Many thanks to Addgenies Ashley Waldron...
  4. Binning Singletons: Tackling Conference Networking When You Don’t Know Anyone

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    Blog Post
    ...keep DNA sequencing singletons (reads that only appear once) in our analysis. Many pipelines toss these...not the more intense mentorship experience that happened at ASM Microbe, we connected people who otherwise...networking and mentoring circles We looked at a lot of approaches scientific meetings have taken to provide mentoring...Shawnna Buttery, Kat Milligan-Myhre, and Pragati Chengappa.  Financial and moral supporters Denise Akob, ...
  5. Controlling for Off-target Effects with a New Genome-wide CRISPR Screen Design

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    Blog Post
    .../Cas9 screens are a high-throughput systematic approach for identifying genes involved in a biological...Cas9 screen design Are non-targeting guides an appropriate control? Many CRISPR screens use non-targeting...Figure 1: Summary of the advantages of the new approach to genome-wide CRISPR screen design.   Bassik...all ~20,500 protein-coding genes. Unique, non-overlapping sites in the genome were targeted and predicted...
  6. Bioinformatics at Addgene

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    Blog Post
    ... expand, we realized the need to modernize our approach to handle larger volumes of data. In this two-...the need to accelerate science using different approaches. We were also encouraged to see that a community...multiple open-source sequence assembly tools at the appropriate place in our workflow without having to translate...computer ran the process. Another design choice we appreciated about Nextflow was how effortless it was for ...
  7. Viral Vectors 101: Preparing Pooled Libraries

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    Blog Post
    .... Transfection of the DNA pooled library and appropriate packaging and envelope plasmids into HEK cells...next step is to make your virus. If the library happens to have a specific protocol for producing virus...protocol on Lentivirus Production provides a general approach for producing lentivirus in Lenti-X 293T cells...don’t hesitate to email us at [email protected]. Happy screening! This blog was written by Addgenie Hannah...
  8. Plasmids 101: Codon usage bias

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    Blog Post
    ...tRNAs correctly, and charge the tRNAs with the appropriate amino acids. If an organism uses only a subset...codons, the ribosome may therefore stall at inappropriate locations or fail to make it through the entire...genes in such a way that their codons are more appropriate for the desired expression host. This is known...ways to encode them and what constitutes the “appropriate” codon is not necessarily obvious. You might ...
  9. Mesothelioma - Causes, Symptoms, and Treatment

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    Blog Post
    ...a danger of getting the disease, even if you do happen to be exposed to asbestos. However, as the U.S....techniques are used as part of a treatment plan, an approach known as multimodal therapy. Sadly, in many cases...the abdominal cavity. This targeted chemotherapy approach has been shown to be successful in killing many...as in the case of Paul Zygielbaum. A different approach to immunotherapy is bevacizumab (Avastin). This...
  10. Using Video to Share Your Science: We Share Science

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    Blog Post
    ...for millions and their visual format is quite appealing to many audiences. While all of these platforms...ever since. The take home message of all of this appears to be that video is the future in terms of communicating...these platforms (such as social networking, visual appeal, instant sharing across multiple other sites), ...private sector researchers, or pretty much anyone applying the scientific method to a question. Videos from...
  11. Data Freedom: The Expansion of Data Sharing in Research Publications

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    Blog Post
    ...detect possible errors in assumptions, use of inappropriate statistical methods, omission of some data, ...distinct (perhaps similarly impoverished lab) might apply different or novel tools and assumptions to reveal...including the generators. This practice doesn’t appear to harm the originating researchers, even if they...given the endless types of data). Rather, PLOS appears to leave the scope of data to be shared to individual...
  12. March for Science

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    Blog Post
    ...Feeling like an apple will fall up instead of down when dropped will not make it happen. In a time when...earlier analogy, no amount of shouting will make an apple fall up. As scientist and politician Rush Holt said... Rush Holt: Lawmakers need to use a scientific approach to formulating views Pew Research Center: Public...
  13. Hot Plasmids: Summer 2024

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    Blog Post
    ...chaperone NPM2, but the technique should be widely applicable to other targets. The modular system of linkers...scFvs for other targets or could be used for applications beyond cryo-EM.   Find plasmids for MagIC-Cryo-EM...plunge freezing represents a convenient, effective approach to protecting your cryo-EM samples from damage...optimizing prime editing systems for specific applications! Delivering prime editors (PEs) via engineered...
  14. Using Phosphoserine to Study Protein Phosphorylation

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    Blog Post
    ...Madison, Wisconsin. It is commonly cited that approximately one-third of cellular proteins are modified ...phosphorylation is far more prevalent than previously appreciated. PhosphoSitePlus, one of the most inclusive databases...protein modification within the context of its appropriate regulation. To avoid these problems, it is also...requires a molecular toolkit containing the OTS, an appropriate bacterial host strain for protein production,...
  15. Plasmids 101: Restriction Cloning

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    Blog Post
    ...toward the cathode (- charge). When the voltage is applied across the gel, the DNA migrates toward the anode...want a “recipient plasmid : insert ratio” of approximately 1:3. Since the number of base pairs for each...should also verify that you are plating on the appropriate antibiotic and try varying the “recipient plasmid...Ideally, once you know that your plasmid has an appropriately sized insert, you should send it for sanger ...
  16. Plasmids 101: Using Transposons in the Lab

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    ...transposition activity  Tol2 transposon plasmids    Applications of transposons Now that you’ve learned about...and Tol2), each of which has several different applications (mutagenesis screens, transgenic animals, gene...Garcia-Perez J (2010) DNA Transposons: Nature and Applications in Genomics. CG 11:115–128. https://doi.org/...Tools: A Review and Guide through Mechanisms and Applications of Sleeping Beauty, piggyBac and Tol2 for Genome...
  17. Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)

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    Blog Post
    ...cells with no Cas9 should appear green and cells with functional Cas9 should appear less green. Note that ...the population, so some of the cells will still appear green despite having active Cas9 (see tip 2). Pro-Tips...by the presence of DNA cleavage products. The appearance of the 428 bp band indicates edits in the BRAF...
  18. Antibodies 101: Flow Cytometry

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    Blog Post
    ...protein composition. It is used for a variety of applications such as measuring target protein expression ...stain the intracellular target. When using this approach, avoid fixation-sensitive fluorochromes such as...assaying a secreted protein, first block the Golgi apparatus with Brefeldin A to inhibit secretion and then...antibodies during the flow cytometry experiment and will appear as a distinct population. After initially gating...
  19. Fluorescent Proteins 101: Photoactivatable Fluorescent Proteins

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    Blog Post
    ...up. Applications of photoactivatable fluorescent proteins Some of the most important applications for ...level of background fluorescence. For any PA-FP application, the brighter the fluorescence the better. More...photostable green and red states. Beyond these imaging applications PA-FPs have been developed for use in yeast,...
  20. Typing CRISPR Systems

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    ...Shakespeare for the first time. The discovery and application of CRISPR-Cas9 kicked off over a decade of frenzied...classification system will help you gain a deeper appreciation and understanding of CRISPR, and perhaps provide...mechanisms in type I-C CRISPR-Cas3 for genome-editing applications. Molecular Cell, 84(3), 463-475.e5. https://...Mashimo, T. (2022b). Genome editing technology and applications with the type I CRISPR system. Gene and Genome...
  21. A Needle in a Base-Stack: Cas9 Structural Biology

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    Blog Post
    ...PAM is so flexible in this state that it doesn’t appear in the crystal structure (Figure 2) at all. When...cleavage. Nucleotides 11-20 are disordered and don’t appear in the Figure 3 crystal structure, reflecting the...which stabilizes the last base pair with an end-capping loop (Nishimasu et al., 2014). This cap is where...with and better understand how to manipulate it. Happy CRISPRing!   Molecular graphics and analyses performed...
  22. Celebrating the 2024 Nobel Prize in Chemistry

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    Blog Post
    ...show the interactions between amino acids in an approximate, two-dimensional map. The Baker Lab wanted to...novel, the principles that created it have been applied to numerous other protein design goals, both from...drugs, all developed with their protein design approach. Congratulations to David Baker, Demis Hassabis...
  23. #DeckTheLab with Tinsel and Test Tubes, Fa La La La La La La La La

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    Blog Post
    ...This edition of the contest is our traditional approach. You might decorate your lab in pipette tip snowflakes...piece of advice - get your PI or lab manager’s approval before letting your imagination run loose on the...diorama. Or maybe you want to create a stack of wrapped gifts using life science catalog pages. Make your...
  24. Editor's Choice, July 2016

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    Blog Post
    ...target and use FRET flurophores for a variety of applications. While editing this post, I learned a lot about...this post to anyone wondering if and how they can apply FRET in their own research. All posts from July ...Blogger Form and we'll be in contact with you ASAP. Happy Reading! Tyler ...
  25. Ghanaian Researchers to Receive Plasmid Awards from Addgene and Seeding Labs

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    Blog Post
    ...concluded, Addgene and Seeding Labs will be opening applications for a second round of plasmid awards on June...excited to discover what new and creative ideas applicants will propose for plasmids from the Addgene community...regions in the near future. Additional Resources Apply to Recieve Equipment through Seeding Labs' Instrumental...
  26. Grad School Advice Part 2: Building Community

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    Blog Post
    ...brains on how they've managed to keep themselves happy during their time in grad school. As you'll learn...student communities 5:59 - 8:16 : What has made you unhappy in grad school? As part of this discussion, we ...to the right that illustrate a grad student's happiness v time throughout graduate school. This is, of...
  27. Plasmid Preps: Different Purity, Different Quantities, Different Uses

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    Blog Post
    ...a plasmid or viral origin. The type of cloning approach you choose ultimately will influence the quantity... for the extraction. Restriction enzyme based approaches can require several hundreds of nanograms or ...assembly or Golden gate assembly use a more direct approach, allowing scientists to clone from a smaller amount...
  28. More Data for You: Find Articles Citing Addgene Plasmids

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    Blog Post
    ...Citing This Plasmid" database - and we always appreciate any help we can get. If you know of an article...PMID) for the article that has used this plasmid. Happy researching! If you need any assistance finding ...have made to the repository, email us and we'd be happy to help. Read More Tips from Addgene Plasmids ...
Showing: 421 - 450 of 954 results