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Showing: 141 - 160 of 895 results

  1. Plasmids 101: Yeast Vectors

    Type
    Blog Post
    Published
    Feb. 25, 2014, 7:11 p.m.
    ...which can be easily maintained when grown on media containing the missing nutrient. Scientists can exploit...the marker gene) For use in: Can this be used in other yeast species? Can this be used for auxotrophic... maintenance within an E. coli cell, but vectors can be widely utilized across many different cell types...those of plants and animals. Unlike bacteria, yeast can post-translationally modify proteins yet they still... chromosome. As the name suggests, these vectors can replicate independently of the yeast chromosome; ...propagate per cell. The copy number of these vectors can also be controlled if specific regulatable elements... yes - Grow with 5-FOA. S. cerevisiae yes - This can complement ura4- S. pombe, but the complementation...

  2. Tips to Make the Most of a Scientific Conference

    Type
    Blog Post
    Published
    May 23, 2019, 12:26 p.m.
    ... scientist whose work you are familiar with. You can tweak your elevator pitch slightly if you know a ...you have a poster or talk at the conference, you can end by asking them to come to your poster or talk...your agenda. Some conferences even have apps you can use to build a schedule. If there is a list of poster...this information on the poster) so your visitors can have a DOI to go to. During the conference Arrive...through the poster.” Have a notebook handy so you can jot down notes or questions from your poster viewers...media. Social media Before the conference Networking can be made easier through social media. Use social media...If you could not attend some of the sessions, you can see if others have live-tweeted as well. In addition...

  3. Making CRISPR Plasmids Using Fragmid

    Type
    Blog Post
    Published
    Aug. 29, 2024, 1:15 p.m.
    ...yourself with a common dilemma. Request the vectors you can find and use them as-is, saving time and effort but...cloning the vectors you actually want. But now you can use the recently developed third option: Fragmid....are the components. Using the handy website, users can simply select the components they want for each module...selection markers. Need fewer components? A spacer can be inserted instead, allowing everything to stay ... individual plasmids and as the Fragmid kit. You can also browse all Fragmid components on the Fragmid...restriction digest, and then plasmid sequencing if you can. To quote Doench, “Whole plasmid sequencing is just...recognition sequence. This means that each component can be designed with an overhang specific to the type...

  4. Fluorescent Proteins 101: Aptamer Fluorophores

    Type
    Blog Post
    Published
    April 11, 2017, 2:30 p.m.
    ...ratios with such techniques can be problematic, however. Well designed aptamers can help reduce such background...temperatures, since proteins can denature while aptamers are more stable and can go through cycles of denaturation...bacteria). That range is possible because aptamers can either fold to incorporate their target molecule,...enzymes, such as: Design and selection for targets can be done largely in vitro; Easy and reproducible ...antibodies for diagnostics know how frustrating it can be to get a “bad batch”); Customizable sensitivity...aptamers glow The flexibility of aptamers means they can be used with or as fluorophores in several different...aptabeacons (or aptatags) were born. Aptabeacons can be created in many different ways, but the general...

  5. Viral Vectors 101: Producing Your rAAV

    Type
    Blog Post
    Published
    July 16, 2024, 1:15 p.m.
    ...viral particles that can transduce cells, which can vary based on target cells and can be altered by freeze... where you can obtain excellent quality, high titer rAAV (including Addgene!), but you can also generate...due to their high density, which can be collected afterwards. You can then remove the density gradient...system, HEK293T cells, expressing Ad E1A and E1B, can be transfected with your transfer plasmid, carrying...titration by qPCR.   Pro tip! An optional media exchange can be performed 12–18 hours post-transfection to help...For certain in vitro and in vivo applications, you can consider using unpurified AAV from the supernatant...Purification To purify your rAAV from your supernatant, you can simply use polyethylene glycol (PEG). We add 25 mL...

  6. Deep Dive: qPCR

    Type
    Blog Post
    Published
    Sept. 29, 2022, 1:15 p.m.
    ...and analysis.  As a rule of thumb, you can assume that qPCR can accurately detect expression differences...differences, and it can even be used to detect a single copy of a transcript. But this sensitivity can sometimes...configurations can help you identify pipetting or evaporation errors. And if time allows, you can fit in a ...statistical significance (Taylor, 2017). With the right conditions and parameters, some people find it can detect... requires significant optimization to ensure the results are correct. Primers and probes can interfere...troublesome, multiplexing your reference gene set can save significant plate space and may be worth the initial...to measure relative quantification of mRNA, which can be used as a proxy measure for differences in protein...

  7. Antibodies 101: Avoiding the Mouse on Mouse Mess in IHC

    Type
    Blog Post
    Published
    Sept. 21, 2023, 1:15 p.m.
    ...in non-specific staining can be a challenge in the best of cases. But it can be particularly challenging...(Figure 2B) Some labs have found that you can significantly decrease background staining by mixing your...same levels of endogenous IgGs, so some researchers can get away with species on species staining with no...    Figure 1: Species on species staining can be a problem because the secondary antibody is not... However, directly conjugated primary antibodies can be expensive and may not provide enough signal to...1998). After washing away unbound fragments, you can then apply your primary and secondary antibodies ... Proper tissue processing and sample preparation can also help reduce the levels of endogenous IgGs in...

  8. Mentoring for Scientists: I Have a Mentor, What Now?

    Type
    Blog Post
    Published
    March 6, 2014, 7:12 p.m.
    ... fun example and you can find it in our Mentoring for Scientists Guide.  You can use it to clarify and...step you can take is to set goals to work on together and track progress at every meeting. It can take many...impart. An undergraduate can mentor a high school student, a graduate student can mentor an undergraduate...Series. The entire series and additional resources can be downloaded in E-Book format at the end of this...setting to your mentoring relationships, so that you can reap rewards in the form of reaching career development...undergraduate, a postdoc can mentor a graduate student – you get the idea. There are many advantages to being a ... Schedule meetings or calls regularly and don’t cancel, so that you drive progress with regular accountability...

  9. Transferable Skills Guide: Creativity

    Type
    Blog Post
    Published
    Jan. 18, 2022, 2:15 p.m.
    ...scientist. In this blog, we’ll delve into strategies you can use to foster innovation. If you identify as a non-creative...yourself a block of time and a quiet space where you can address the issue head on. Prepare for this time ...environment where everyone has their ideas heard.  So how can we assure that all participants feel comfortable ...devil’s advocate is elected, others may feel they can share opposing opinions as well. I am sure we have...goal is to generate unique ideas. Recognize ideas can come any time. Meetings shouldn’t be the only place...ideas are not well received, understand that people can be resistant to change. Ask your peers or your superiors...fixated with the idea of making something the best it can be that it seems impossible to start at all. This...

  10. 10 Steps to a Perfect Science Talk

    Type
    Blog Post
    Published
    Aug. 23, 2016, 2:30 p.m.
    ...the best speaker you can be. Seek out opportunities to practice and over time you can master this important...solutions—public speaking is skill that any scientist can learn.  Any time you give a science talk, you are... added bonus to knowing your audience is that it can help you avoid making mistakes. I once made a joke...a narrative connecting cause and effect. Stories can and should be very simple. The goal is to get the...to include that material, but don’t force it. You can always relate to scientists by explaining how your... real audience Good, even great, speaking skills can be attained by almost anyone with enough practice...previous year. Usually fewer than 10% have done so. How can you expect to deliver a great public presentation...

  11. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    Published
    June 28, 2016, 10:59 a.m.
    ...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...

  12. Thoughts on the Future of Research 2015 Boston Symposium

    Type
    Blog Post
    Published
    Nov. 25, 2015, 4 p.m.
    ...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 ...

  13. Viral Vectors 101: Inverted Terminal Repeats

    Type
    Blog Post
    Published
    Oct. 19, 2023, 1:15 p.m.
    ...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...

  14. 6 Steps to Submitting a Resume That Gets Seen

    Type
    Blog Post
    Published
    July 9, 2015, 2:30 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    June 8, 2017, 2:30 p.m.
    ...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...

  16. Viral Vectors 101: Virus Safety

    Type
    Blog Post
    Published
    June 22, 2023, 1:15 p.m.
    ...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...

  17. To Each HIS Own

    Type
    Blog Post
    Published
    March 2, 2023, 2:15 p.m.
    ...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...

  18. 10 Basic tips for mammalian cell culture

    Type
    Blog Post
    Published
    July 12, 2018, 1:09 p.m.
    ...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...

  19. Antibodies 101: Flow Compensation

    Type
    Blog Post
    Published
    May 16, 2024, 1:15 p.m.
    ...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...

  20. Why Do I Need an MTA Anyway?

    Type
    Blog Post
    Published
    May 1, 2014, 8:23 p.m.
    ...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...
Showing: 141 - 160 of 895 results