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Showing: 281 - 300 of 1075 results

  1. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    ...are looking, you can either scan the genome yourself for potential gRNA sites, or you can utilise several... or you can order them yourself through whatever DNA supply company you currently use. You can also find...sequencing comes back. Alternatively, if you can afford to wait, you can hold off on genotyping until the pups...targets in that region. The CRISPR Software Matchmaker can be used to determine the pros and cons of using many...genomic fragment containing the sgRNA target sequence can be placed in multi-cloning site (MCS) between EGFP... and sgRNA expression cassette. The target sgRNA can be cloned directionally into the BbsI site. (c) The...sequence surrounding your gRNA. Once amplified, you can insert this region into a pCAG-EGXXFP plasmid using...

  2. Thoughts on the Future of Research 2015 Boston Symposium

    Type
    Blog Post
    ...out to your peers and other mentors who can help you. You Can Talk to Your Editors I wish I had been... was “How many graduate students/post-docs?” You can find some of the more detailed questions the organizers...chosen career path,” but individual institutions can have many different names for the positions commonly...universities have simple postdoc numbers sorted out there can be gaps in knowledge of demographics and the information...environments. Slide from Paula Stephan’s Talk (All slides can be found here). Importantly, the panel offered some... panel, focused on showing researchers that they can take steps to achieve success in their careers. The...the coming months and welcome guest bloggers who can provide additional advice to trainees navigating ...

  3. Viral Vectors 101: Inverted Terminal Repeats

    Type
    Blog Post
    ...sequences determines the orientation of the ITR, which can either be “flip” or “flop” (Figure 2). Additionally... elements of AAV and rAAV vectors, modifications can drastically affect gene transfer efficiency and transgene...important for viral genome replication, these mutations can affect transduction efficiency and subsequent transgene...sequence containing transcription factor binding sites can increase transgene expression and transduction efficiency...efficiency (Shitik et al. 2023). Transgene expression can be altered by using ITR sequences from different ...activity (Earley et al. 2020), but non-AAV2 ITRs can be used to drive gene expression at varying capacities...capacities. AAV vectors containing the AAV2 ITRs can be cross-packaged with capsids from multiple AAV serotypes...

  4. 6 Steps to Submitting a Resume That Gets Seen

    Type
    Blog Post
    ...that can happen is someone will say no, but if you don't ask, you will never get the help that can make...the most effective tactic a scientist in training can adopt to ensure opportunities in the future. Studies...relationships as part of the job application process can dramatically increase the chances of being seriously... submitted The structure and content of a resume can be crucial for a successful application. It takes...good at learning new things—this is a strength you can draw on and demonstrate in your application). Beware...During the entire process, use whatever means you can to get your resume into the hands of the hiring manager...great way to find an appropriate liaison, but you can use alumni databases, friends, and even relatives...

  5. Choosing the B(right)est Fluorescent Protein: Photostability

    Type
    Blog Post
    ...probes. You can follow him on twitter: @joachimgoedhart.   Marieke Mastop is a PhD candidate at the Section... best fluorescent proteins for live cell imaging can be excited many times, thereby producing a large ...power is a property of the fluorescent protein. This can differ between fluorescent proteins in the same spectral..., the photostability of the fluorescent proteins can be directly compared. An example of a photostability...distribution. Therefore, photostability measurements can be performed with soluble fluorescent proteins or...make these measurements and hope this information can help guide your understanding of reported data. Photobleaching...loss of fluorescence, which, in its simplest form, can be described by a mono-exponential decay (analogous...

  6. Viral Vectors 101: Virus Safety

    Type
    Blog Post
    ...needed, a layer can quickly be removed and disposed of without risking exposure. You can request disposable...meaning it can’t replicate and continue the infection cycle once inside the host. While the virus can still... to integrate into proto-oncogenes, which can lead to cancer. These viruses need to be handled in a BSL...useful because they’re derived from viruses that can infect people, cells, and animals. Some of these ...these viruses are completely inert, but others can cause diseases. Nervous about handling viruses? That’s ...to take, and if something does happen, ensure you can receive appropriate treatment and/or monitoring. ...as part of the viral infection process (though it can integrate at chromosome 19q13.4 qtr. (AAVS1)). For...

  7. To Each HIS Own

    Type
    Blog Post
    ...examination of specific proteins within a system. This can be pretty complicated given that a single cell has... purification. In this strategy a single protein can be isolated from a complex mix using affinity chromatography... tag, on the protein. While a variety of methods can be used to successfully isolate proteins, this blog...metal ions such as Ni2+ or Co2+ (Hochuli 1988), and can elute proteins with up to 95% purity (Janknecht 1991...histidines are present in most production systems and can bind and co-elute with the protein of interest (Bornhorst...denaturants (Hochuli, 1988). In some cases, proteins can be bound under denaturing conditions to expose the...preferable over the other however, tag placement can affect protein function. B, The string of histidines...

  8. 10 Basic tips for mammalian cell culture

    Type
    Blog Post
    ...stock, while you can use the remaining vial to grow and freeze additional cells that can be used as your...UV light 3-4 times a week for up to half an hour can help keep your hood contamination free, but you should...disinfectant (most people use 70% ethanol, but you can also use Incidin wipes) before and after your work...materials you’ll need to bring into the hood and can wipe them all down before you start. This will also... difficult to clean your workspace properly, but can also disrupt the laminar air flow around your work...have a light microscope close to the hood so you can view your cells regularly (both before entering the... you’re working with so that cross-contamination can be identified. Even if some mammalian cells lines...

  9. Antibodies 101: Flow Compensation

    Type
    Blog Post
    ...markers can be challenging! But we hope this guide can help you get started. Remember, you can always ...This can be helpful as you're learning! Ashlyn has a research background in CD8+ T cells and cancer immunotherapies...By compensating between fluorescent channels, you can correct for this spillover and more easily separate...your samples. This means that, unlike voltages, you can adjust compensation values later in your analysis...for APC (Figure 1B). For single-color samples, you can only change the compensation values for whichever....     In general, most spillover between samples can be fixed with fairly low compensation values (0-50...machines or the program you’re using, but usually you can click on the correct box and either manually change...

  10. Why Do I Need an MTA Anyway?

    Type
    Blog Post
    ...soul of Addgene’s mission), but legal complexities can arise when scientists are sharing their materials...publications, and commercial use often come up. An MTA can resolve those questions! MTAs protect you and your...all those benefits, we know that legal documents can get tricky. Here’s how Addgene helps: For Providers...distribute your materials under the UBMTA terms, so you can rest easy knowing your rights – and the rights of...generates the MTA and emails it straight to you, or we can send it directly to your institution’s MTA coordinator...Organizations that have opted into our eMTA system can even sign electronically or choose to auto-approve...result in quick approvals, so your plasmid requests can get to your lab faster. For Everybody: Friendly...

  11. Save Time and Money by Making Your Own Competent Cells

    Type
    Blog Post
    ...performed at Addgene, this adds up to a significant time savings. You can also save yourself time if you're ...step. Read on to find protocols and tips that you can use in your own lab. How to make your own competent...competent cells because cells prepared using this kit can be transformed without heat shock. Detailed protocols...cells using the protocols provided above, then you can find the subsequent transformation protocols below... Cells prepared using the Z-competent TM buffer can be easily transformed without a heat-shock step. ...resistance gene time to be expressed.  However, you can skip this incubation step and go straight to plating...other competent cells) to ensure that your cells can be readily transformed. Compare the number of colonies...

  12. ReFigure: Save Scientific Figures into Dashboards and Share Your Insights

    Type
    Blog Post
    ... reach. We can’t save you the hard work of reading papers and doing experiments but we can definitely ...large section of open access life sciences data. You can use ReFigure to coherently save analyses with the...insights on a topic. Beyond collating other’s data, you can even include your own data in a ReFigure. For example...Figshare is currently supported by ReFigure beta) you can include it in a ReFigure. ReFigures are good for ...proposals. ReFigures are one such research product which can convince reviewers of your expertise and insight....minutes, visit our YouTube Channel. Currently we can only connect figures from PubMed Central, eLife, ...Download ReFigure extension and visit ReFigure.org. You can reach the ReFigure team at [email protected] ...

  13. IBEX Knowledge-Base: A Data Resource for Multiplex Tissue Imaging

    Type
    Blog Post
    ..., 2020, 2022). I do mean highly multiplexed: you can image over 65 different parameters using this method... from Radtke et al., 2020). This flexible system can be used with many conventional microscopes, amplified...that they used in their experiments. Researchers can then visit the IBEX imaging community discussion ...conversations with other users. The entire dataset can be downloaded from Zenodo, including the reagent ...for easy filtering and sorting. Reference images can be viewed by clicking on the links in the "Agree"...detergent, antigen retrieval conditions, and more. Users can also recommend or not recommend a particular reagent...discussion forum where scientists from around the world can ask and answer questions. Andrea and Ziv were...

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

    Type
    Blog Post
    ...Addgene’s customer service philosophy Customer service can be a hassle. You finally make it through a 20-step...this is why we have a dedicated support team that can help you through any search, ordering, technical,...adding self service to the repertoire of tools you can use to get support for our resources with our new...asked questions. On the Help Center webpage, you can browse through our various FAQ categories or search...enter your name and email address, after which you can describe your issue and press send. Doing so will...directly to our support team’s attention, where we can then offer the same high level of direct service ...from us.   Try out the Help Center! The Help Center can be found at help.addgene.org, and the widget is on...

  15. Advice for Choosing a Research Project

    Type
    Blog Post
    ...similar interests and goals. The following questions can serve as a guide when selecting a research project...What are possible pitfalls in your strategy and how can you mitigate them? If you were to write a grant for...skills do you need to use to reach these milestones? Can you get assistance from people who have the required...What is your timeline? A typical research project can take several years to complete and has multiple benchmarks... new tools and methods that have been developed. Can you think of a project that combines an exciting ...own subject - maybe something used in astrophysics can be applied to fungal biology.” Connie Cepko advises...complexity of biology and break apart a piece that can address a topic in a mechanistic way using the technology...

  16. Trading Spaces - Visiting Scientist Edition

    Type
    Blog Post
    ... with guidance and training so that you yourself can become an expert and bring the knowledge back to ... perspectives. For trainee level researchers, it can expose them to a new institute and mentor that they...experiencing a new city or maybe even a new country can also be exciting! Identifying a host institute or...personally know or your lab collaborates with that can fulfill your research needs, that is probably the...visiting positions within defined labs which you can apply to. These structured types of placements often...visit, if you don’t have a formal connection, you can just reach out to start the conversation. Many researchers...plan with your host prior to arrival so that you can work together to ensure your expectations are reasonable...

  17. What Good Citizenship Can Do for Reproducibility in Science

    Type
    Blog Post
    ...before their paper can be published. To a scientist, all of this checking and formatting can feel like yet ...lot of time. It doesn’t have to be this way. We can never completely eliminate the risk that a minor ...stability in an incubator), but there’s a lot you can do to help other scientists both what you did and...scientific citizen” and making sure you do what you can to help your fellow researchers advance science overall... Communication is the issue Much of the problem can be attributed to communication, and in particular...Admittedly, the differences are often very subtle, and it can take a lot of detective work to figure them out. ...accessible and consistent way so other researchers can use them, to making sure that methods and protocols...

  18. Summer SciComm: Preprints

    Type
    Blog Post
    ...Preprints can be read freely by anyone and are typically free of charge for authors to post. They can be assessed...materials available to the community significantly sooner. Plus, authors can include the Addgene ID for their...them as best you can! At the time of writing, Mike Lacy was also employed by the American Society for Cell...sharing results faster and more broadly, scientists can receive recognition for their efforts, inspire future... preprint as evidence of their productivity that can be shared with hiring and review committees, as opposed...comments, reviewers’ comments, and author responses can be shared directly alongside the preprint while the...awaiting a final publication in a journal.  Preprints can be one tool in a combination of strategies for your...

  19. Viral Vectors 101: Viral Applications

    Type
    Blog Post
    ...Viruses are the SUVs of molecular biology – they can deliver materials to specific destinations, haul ...research all the way to clinical gene therapy. But how can you use these amazing tools in your experiments? ...cargo is typically their own genome, but this step can be engineered to be anything you want delivered to... to your target cells. This delivered DNA/RNA can then be either permanently integrated into the genome...application, our Viruses as Biological Tools blog can help you learn about the different types of viruses...degradation. Viral introduction of shRNA plasmids can induce gene-specific silencing temporarily, long-...turning expression off permanently. Viral vectors can also be used to express proteins. Viral expression...

  20. The time and cost required to make a plasmid

    Type
    Blog Post
    ... way, you can run one PCR, one gel, etc. for multiple plasmids at once. Over time, this can save you a...how much time you have to spend cloning before you can start your experiment? What about all the reagents...eternity of cloning, waiting, and repeating before you can finally dive deep into generating the data you’re...Table 1). Restriction enzymes and vector purchases can raise the cost anywhere from a few dollars to over...varies hugely based on a number of factors, but we can assume it’s at least equal to the $155 spent on reagents...short on money or time, there are a few options you can explore to maximize the use of both when making plasmids...) Ask the PI If you want a plasmid for free, you can always reach out to the PI or corresponding author...
Showing: 281 - 300 of 1075 results