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Showing: 401 - 450 of 954 results

  1. Pairing CombiGEM and CRISPR for Combinatorial Genetic Screening

    Type
    Blog Post
    ...high-throughput screening are vastly useful for mapping the combinatorial genetics that underlie complex...research questions. CombiGEM has been successfully applied to functionally characterize combinatorial gene...realization of a variety of perturbations and applications in functional genomics, cell reprogramming, ...He is particularly interested in developing and applying new technologies to interrogate and understand...

  2. Getting Started with Behavioral Neuroscience: Standardizing Experiments and Using Viral Vectors

    Type
    Blog Post
    ...need to be given some time to adapt to a testing apparatus. General stress levels of the test animal may ...the humans’ audible spectrum. The experimental apparatus should be cleaned thoroughly between animals as...excitation or inhibition upon light (or drug) application.  Genetic manipulation can also reduce expression...after injection. Therefore, it’s crucial to appropriately time the injection about two weeks before the...

  3. Engaging with science and society at pgEd

    Type
    Blog Post
    ...these tough issues is essential for building a rapport with these communities, which is in turn crucial...latest developments in genetics research, their applications and regulatory policies; keeping tabs on the...scientist. After all, scientific research does not happen in a vacuum that is isolated from the rest of society...and who will be affected by the knowledge and applications generated from this research. At both a practical...

  4. Adenoviral Vector Production and Troubleshooting

    Type
    Blog Post
    ...vectors (AdV) are attractive vectors for research applications and gene therapy: they can be produced at high...AdVs is what makes them ideal candidates for applications in oncolysis and vaccination!) In the 65 years...Adenovirus-mediated gene delivery: Potential applications for gene and cell-based therapies in the new...

  5. Corralling and Cornering the Goldilocks Antibody

    Type
    Blog Post
    ...differently – happens when the antibody is produced from a new set of animals), and wrong application (different...the best, or even which one will work for your application? Like many scientists, I have spent hours sifting...differently over time), compatible with the desired applications (such as immunohistochemistry (IHC) or western...

  6. Viral Vectors 101: Calcium Sensors

    Type
    Blog Post
    ...Additional red sensors based on mRuby (jRCaMP1) and mApple (jRGECO1a) are also available. Detection methods...to increased intracellular calcium. It has been applied to mark active populations of neurons in vivo to...expression and downstream monitoring. This targeted approach allows scientists to focus on specific cell populations...Vectors 101: Biosensors Viral Vectors 101: Viral Applications Viral Vectors 101: An Introduction to AAV ...

  7. Viral Vectors 101: Chemogenetics

    Type
    Blog Post
    ... the end goal, it is best to work with FDA/EMA approved drugs so that your results are viable for translation... be mitigated by using low doses and running appropriate controls (Jendryka et al., 2019), they have resulted...brain and some, such as varenicline, are already approved for clinical use. Varenicline has been further...chemogenetics for research and potential clinical applications. Science 6436: eaav5282. https://science.sciencemag.org...

  8. A Look at Addgene's QC Process

    Type
    Blog Post
    ...mutated; elements that are difficult to sequence; approaches to confirm plasmid sequences without a reference...program called Geneious. Here, the raw reads are mapped against a reference sequence, if one was provided... determine if there are any notable QC issues. Mapping the raw NGS reads to a reference sequence can also...confirm their presence with confidence. If this approach does not resolve the identified differences, the...

  9. How-to: 5 Steps to a Great Panel Discussion

    Type
    Blog Post
    ...organizers and moderator should have a well-planned approach to making the event interesting and engaging. ...information and remind them about the event as it approaches. Room set-up and logistics To make a really...and forth within the panel. Go with it when this happens, but get things back on track if the direction ...talk and the speaker goes over 1 hour, watch what happens to the attention of the audience – phones come ...

  10. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

    Type
    Blog Post
    ...have gotten a lot of press for being a promising approach for treatment or prophylaxis of COVID-19. Researchers...nanobodies that bind to coronavirus spike proteins (Wrapp et al., 2020), which sit on the surface of coronaviruses...BioRxiv.  https://doi.org/10.1101/2020.04.16.045419.  Wrapp D, De Vlieger D, Corbett KS, Torres GM, Wang N, ...single domain antibodies for the conformational trapping of membrane proteins. eLife 7: . https://doi.org...

  11. Plasmids 101: Protein Expression

    Type
    Blog Post
    ... experimental, biotechnological, and medical applications. Researches can visualize proteins in vivo by...protein you need   Your plans for downstream applications In this blog post we will summarize some of...proteins as bacterial cells are not equipped to add appropriate post-translational modifications. In addition...acids that are useful in different downstream applications. Cell-free expression systems however, are expensive...

  12. Twenty Years of Addgene Sharing: CRISPR

    Type
    Blog Post
    ... per year, or approximately 104 plasmids shipped per working day, including appropriate MTAs and customs...of CRISPR tools has grown steadily, gaining approximately 2,000 plasmids/year from 2017 to 2023. This ... joining the Addgene CRISPR community, with approximately 100 new CRISPR depositors each year.    ...

  13. Antibodies 101: Conventional vs Spectral Flow Cytometry

    Type
    Blog Post
    ...order to correct for this, compensation has to be applied to “subtract” this spillover and correctly distinguish...designing your panel. If you have two highly overlapping cellular markers, it is important to pair them...have a similarity index of 0.87; therefore, two overlapping cellular markers should not be paired with these...index of 0, which would be a better choice for overlapping markers. Compensation/reference control(s) Compensation...

  14. The AAVantages of AAV in CRISPR Screens

    Type
    Blog Post
    ...working with mice and slightly less red tape for approval processes. Additionally, while both AAV and lentivirus...have capacities of 8–12 kb, while AAV only has approximately 4.7 kb — which means that when using Cas9, you... Platforming AAV Interest in interrogating and mapping the genetic landscape of tissue-specific diseases...Performing In Vivo CRISPR Screens Using the FITS Approach Additional resources at addgene.org CRISPR pooled...

  15. Experimenting with New Careers while in Grad School

    Type
    Blog Post
    ...months, and we got one submission. I was a bit disappointed in the lack of participation, but I decided ... passed these dinosaurs every day. They made me happy. Grad school can provide plenty of negativity, so... so I volunteered at a place that made me happy. I started volunteering in the “Dinosphere” almost every...was worried about becoming one of those angry, disappointed graduate students because I wasn’t sure what...

  16. INTEGRATE: Bacterial Genome Engineering Using CRISPR-Transposons

    Type
    Blog Post
    ... and have been adapted for a wide variety of applications - as mutagenesis tools for genetic studies (...Recently, the lab introduced a streamlined expression approach (Fig. 2A), involving two-plasmid or single-plasmid...at one or multiple desired genomic loci. Many applications of these systems have already been demonstrated...C , Xu M, Zhou Z, Smock DC, Tang K, Owens TK, Krishnappa N, Sachdeva R, Deutschbauer AM, Banfield JF, ...

  17. Great Results Start with Great Standard Curves

    Type
    Blog Post
    ... critical for a number of popular scientific applications, and the quality of a standard curve can make... then generate a trendline for the data. The appropriate trendline varies between assays and may be linear...for an acceptable R2 but for many scientific applications users aim for an R2 above 0.95.    Additional... from decreased specificity, especially as it approaches the minimum and maximum thresholds. If the measurements...

  18. CRISPR 101: Making a Knock-In Cell Line

    Type
    Blog Post
    ...cells stay in an HDR-promoted cycle longer. This approach may have some negative outcomes on viability if... as 10 hours, so read up on the timing of this approach carefully (Lin et al., eLife)! Use a Cas that...facilitates HR What if promoting HR was as simple as swapping out your Cas9? It can be! Cas12a is a Cas family...knock your knock-in experiment out of the park. Happy editing!   Resources and References References...

  19. Introducing Addgene's New Search - Find Plasmids More Easily For Your Research!

    Type
    Blog Post
    ...down to the exact plasmid you need. The ability to apply a wide range of filters isn’t new to Addgene’s plasmid... it’s also a convenient most requested filter. Applying a second filter, this one from the ‘Requests’ ...consider. If you get this far and realize you need to apply different filters you do not need to start a new...” plasmids returns more than 1300 results, and applying a filter for “CRISPR” plasmids keeps more than...

  20. New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries

    Type
    Blog Post
    ...activity. In this study scientists examined approximately 1,900 sgRNAs and identified sequence features...substantially identified more hits than the SAM library approach in a comparison study screening for vemurafenib...ORF overexpression library identified several overlapping genes, most genes were unique to the individual...Calabrese pooled library and lentiviral prep here! Applications for the Root and Doench CRISPR libraries Overall...

  21. Minigenomes - a Safe Way to Study Dangerous Viruses Like the Ebola Virus

    Type
    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...

  22. Your Lentiviral Plasmid FAQs Answered

    Type
    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...

  23. Technical Design of a Western Blot

    Type
    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...

  24. Controlling for Off-target Effects with a New Genome-wide CRISPR Screen Design

    Type
    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...

  25. Bioinformatics at Addgene

    Type
    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 ...

  26. Viral Vectors 101: Preparing Pooled Libraries

    Type
    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...

  27. Binning Singletons: Tackling Conference Networking When You Don’t Know Anyone

    Type
    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, ...

  28. Plasmids 101: Codon usage bias

    Type
    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 ...

  29. Mesothelioma - Causes, Symptoms, and Treatment

    Type
    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...

  30. Using Video to Share Your Science: We Share Science

    Type
    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...

  31. Data Freedom: The Expansion of Data Sharing in Research Publications

    Type
    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...

  32. March for Science

    Type
    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...

  33. Hot Plasmids: Summer 2024

    Type
    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...

  34. Using Phosphoserine to Study Protein Phosphorylation

    Type
    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,...

  35. Plasmids 101: Restriction Cloning

    Type
    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 ...

  36. Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)

    Type
    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...

  37. Plasmids 101: Using Transposons in the Lab

    Type
    Blog Post
    ...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...

  38. Fluorescent Proteins 101: Photoactivatable Fluorescent Proteins

    Type
    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,...

  39. Antibodies 101: Flow Cytometry

    Type
    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...

  40. Typing CRISPR Systems

    Type
    Blog Post
    ...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...

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

    Type
    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...

  43. Celebrating the 2024 Nobel Prize in Chemistry

    Type
    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...

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

    Type
    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...

  45. Editor's Choice, July 2016

    Type
    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 ...

  46. Ghanaian Researchers to Receive Plasmid Awards from Addgene and Seeding Labs

    Type
    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...

  47. Grad School Advice Part 2: Building Community

    Type
    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...

  48. Plasmid Preps: Different Purity, Different Quantities, Different Uses

    Type
    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...

  49. More Data for You: Find Articles Citing Addgene Plasmids

    Type
    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: 401 - 450 of 954 results