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We narrowed to 936 results for: TIM;

Showing: 421 - 440 of 936 results

  1. Viral Vectors 101: Producing Your rAAV

    Type
    Blog Post
    ...crude cell lysate (Benyamini et al., 2023) to save time and cost.  Purification To purify your rAAV from...dose (TCID50) (Zen et al., 2004).  To choose the optimal titering assay, you’ll need to understand how different...non-genome-containing (empty) capsids, thus leading to an overestimate of transducible units in the virus preparation...Ehrhardt, A., & Baiker, A. (2012). Universal real-time PCR for the detection and quantification of adeno-associated...

  2. PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs

    Type
    Blog Post
    ...experiments, thus saving you time, sweat, and money on extensive optimization efforts. This holds true for...score ranking (Kim et al., 2019). PRIDICT saves time and resources by selecting the most promising pegRNA...Koeppel et al., 2023), their predictive power is sometimes limited across different targets due to variations...

  3. Pooled Library Amplifications

    Type
    Blog Post
    ... 200x coverage, meaning that you need 30 to 200 times as many colonies as individual plasmids in your ...colonies, you’ll likely rely on serial dilutions to estimate transformation efficiency (Figure 2).    ... Figure 2: Example of a serial dilution to estimate transformation efficiency. Figure used with permission...temperature (e.g., 30 °C), and limiting your growth time (e.g., 16-18 hours). Finding reference protocols...

  4. Plasmids 101: 5 factors to help you choose the right cloning method

    Type
    Blog Post
    ...cloner, you’re likely not intimidated by having to pick the right enzymes, optimize reagent concentration,... cloning is another good option to consider when time is of the essence. This method requires a vector...two to generate templates for the cloning process. Time spent actually doing the experiments are also generally...

  5. 10 Basic tips for mammalian cell culture

    Type
    Blog Post
    ...nothing is covering it. Turning on the UV light 3-4 times a week for up to half an hour can help keep your...stop growing (contact inhibition) and it takes them time to recover after they are passaged. This could change...borrowing’ cells that have already been passaged many times and could be contaminated.Learn how Addgene uses... and morphology at high passage numbers (>30-40 times), depending on the cell line so old cells should...

  6. Hot Plasmids and Viral Preps - May 2021

    Type
    Blog Post
    ...library and characterized them by ELISA, homogeneous time resolved fluorescence, and suspension bead assay...degradation, this protein compartment was used to optimize D-glucaric acid biosynthesis. This MPyV system...way to use the accumulation of indels to measure time. These plasmids with different promoters driving...constructs. zCas9i is a high efficiency intron-optimized SpCas9-coding gene. SlugCas9 is a compact Cas9...

  7. Experimenting with New Careers while in Grad School

    Type
    Blog Post
    ... to them about my interests and my ideas all the time. Once, I had this crazy idea that I needed help ...and departments Graduate school was an excellent time to try new things, develop new skills, and meet ...from the experts in science communication in real time! I follow great online science communicators like... why I still love science. Keep building on your time in graduate school I graduated in the fall of 2015...

  8. Anatomy of a Plasmid Page at Addgene

    Type
    Blog Post
    ...been ordered more than 20 times; red, more than 50; and blue, more than 100 times! When you search for plasmids... plasmids which have been ordered more than 100 times).     I guess it's cool to be popular, but how ... number (if known), antibiotic resistance, and optimal growth temperature are listed, as well as the name...

  10. Course-Based Undergraduate Research Experiences (CUREs)

    Type
    Blog Post
    ... teaching responsibilities left him with limited time for one-on-one mentoring. His solution? Bringing...particularly amenable to in-class labs. Provost ultimately settled on a two-semester model, in which students...Maximizes Reach Many new professors are wary of the time and effort and/or perceived professional risk involved...try implementing one. It's likely only a matter of time before a CURE trainee goes on to win a Nobel!   ...

  11. A Career in Grants: How To Become a Grant Professional

    Type
    Blog Post
    ...grant professionals have similar stories about a time when their passion for an organization or a program...area of expertise. Whether you’re seeking a full-time job or want to be an independent grant consultant...and grant management.  Your niche will change over time. Going into the grants profession with a concept... will save you and the organization(s) you serve time and resources. 3. Get a mentor, find your people...

  12. Tips for Writing a Good Cover Letter

    Type
    Blog Post
    ...they were ten years ago, and it does take a bit of time to craft one. We still think they're useful, so ...applying to the specific company or lab. Take some time to research the lab or company via their papers ... to start from a standard version, but take some time to make it fit the job you’re applying for. The ...professional. Always review grammar and spelling several times and ask someone else read your cover letter (as ...

  13. Transferable Skills Guide: External Collaborations

    Type
    Blog Post
    ...software development resources into it. At the same time, this project gives Snapgene great exposure to the... part of my work. However, this is not the first time I’ve had a role in organizing collaborations. During...companies and then convince the contacts to give me some time to talk or to at least read my emails. Then pitching...Personally, I find this to be the most challenging and time-intensive part of relationship building. However...

  14. Antibodies 101: Immunoprecipitation

    Type
    Blog Post
    ...However, the two different approaches allow for optimized strategies for differing amounts of target proteins...following the incubation step. The wash buffer has an optimal pH and ionic strength that breaks weak interactions...antibody intact. The wash step is repeated several times to ensure non-specific proteins are completely removed...isotype control antibody. Alternative applications Oftentimes, an IP-appropriate antibody simply isn’t available...

  15. Plasmids 101: Plasmid Incompatibility

    Type
    Blog Post
    ...during cell division simply does not allow for enough time for correct partitioning to occur (Diaz et al., ... low copy plasmid is more likely to be lost over time.  Is plasmid incompatibility going to affect my ...clarity, a wet lab transformation experiment will ultimately inform whether your plasmids are compatible or...beginning to use plasmid incompatibility to combat antimicrobial resistance. Using novel plasmids based partly...

  16. AAV Titers: Where Do They Come From And What Do They Mean?

    Type
    Blog Post
    ...efficiencies (Wang et al., 2013). Although you can optimize your primers for each individual sample, this ...ensure accurate and reliable results, we have optimized our assay in a few ways:  Absolute quantification...titrating each lot of AAV in vivo to determine the optimal dosage. At Addgene, we don’t reserve lots, but ...quantification by droplet digital PCR versus analog real-time PCR. Nat Methods. 2013 Oct;10(10):1003-5. PubMed...

  17. Plasmids 101: The Promoter Region – Let's Go!

    Type
    Blog Post
    ...gotten to know what a plasmid is in general, it's time to take a look at some of its parts. Elements such...promoters are constitutively active and on all the time while others are more carefully controlled. Regulated...might act only in certain tissues or at certain times in development or there may be ways to turn them...promoters available to scientists. Plasmids are oftentimes put to therapeutic uses, and in those cases it's...

  18. Viral Vectors 101: Integrase-Deficient Lentiviral Vectors

    Type
    Blog Post
    ...vectors can be produced easily, without the need for time-consuming purification processes, and are powerful...integrate into the host cell’s genome. However, sometimes inte-great-ion isn’t so great. In this post, we...expression of gene products (Wu et al., 2004). Over time, the expression diminishes as the episomes are lost...express a protein or nucleic acid for only a limited time, IDLVs are ideal. As the episomes are lost during...

  19. March for Science

    Type
    Blog Post
    ...hold a doctorate degree, are currently working full-time, and have received a research grant in the last ... down when dropped will not make it happen. In a time when a top official can be quoted as saying “I’m...primary contributor' to global warming The New York Times: With Trump in Charge, Climate Change References...Technology: Crossing The Imaginary Line The New York Times: A Scientists’ March on Washington Is a Bad Idea...

  20. Harnessing Bacterial Toxins for Allelic Exchange

    Type
    Blog Post
    ...cassette. This leaves a “scar” in the chromosome, sometimes interfering with expression of surrounding genes...technique, and is less prone to off-target mutations. Ultimately, successful allelic exchange requires two sequential...mutants, but it frequently requires considerable optimization to ensure strong counter-selection. Anecdotally...magenta. This is particularly valuable when first optimizing conjugation in strains that don’t conjugate that...
Showing: 421 - 440 of 936 results