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Showing: 421 - 440 of 923 results
  1. Anatomy of a Plasmid Page at Addgene

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    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...
  2. Course-Based Undergraduate Research Experiences (CUREs)

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

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

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    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...
  5. Transferable Skills Guide: External Collaborations

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    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...
  6. Tips for Writing a Good Cover Letter

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    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 ...
  7. Antibodies 101: Immunoprecipitation

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    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...
  8. Plasmids 101: Plasmid Incompatibility

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

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    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...
  10. Multicolor Animals: Using Fluorescent Proteins to Understand Single Cell Behavior

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    Blog Post
    ...Each cell remains the same color for its entire lifetime and daughter cells retain the same color, allowing...allowing for the fate mapping of cell populations over time. The ability to track single cell dynamics at the...to create foci of proliferating cells. Tetbow optimizes Brainbow for current technologies Tetbow (Sakaguchi...that incorporates chemical tags instead of XFP to optimize it for tissue clearing, a method that makes tissues...
  11. March for Science

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    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...
  12. Harnessing Bacterial Toxins for Allelic Exchange

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    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...
  13. Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus

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    Blog Post
    ...and this is likely a rate limiting step, but optimization of the exon-splitting site sequence can enhance...and acceptor sequences were tested and with the optimal sequences, the group achieved ~80% of muscle cells...level of transgene expression, and may need to be optimized for each unique application (Trapani et al., 2015...resources on the Addgene blog: Find tips for a first time AAV user Learn about how AAV is used in retinal ...
  14. Proximity Labeling: A Powerful Tool for Protein Complex Purification and Proteomic Mapping

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    Blog Post
    ...Addgene! Since the development of BioID and APEX, optimized versions of BioID and APEX have been developed...promiscuous biotin ligase The Roux Lab in 2016 optimized BioID to create BioID2, a smaller and improved...hypothesized that the low sensitivity was due to suboptimal folding or stability. Thus they conducted multiple... significantly greater than miniTurbo at earlier time points but miniTurbo exhibits less labeling prior...
  15. Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

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    Blog Post
    ... and Lichtman estimate that Brainbow’s colorful labeling has decreased the mapping time for a given section... were limited due to low resolution or were very time-intensive, necessitating the development of a new...is truly fitting for this colorful technology. Optimizing the system: Brainbow-3 Although Brainbow provided...
  16. Recombinase-based State Machines Enable Order-dependent Logic in vivo

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    Blog Post
    ...behaviors, but little is known about the order or timing of these factors. Recombinase-based state machines...reaction can only be performed on one register at a time. If a particular application makes it difficult ...machine design space is highly degenerate and often times there are multiple register designs that may be ...? We tested a two-input (ATc and Ara) RSM over a time series of increasing input exposure lengths and ...
  17. Sequencing Options for CRISPR Genotyping

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    Blog Post
    ...2015). Meanwhile, Sanger sequencing is laborious, time consuming and cannot be applied to heterogenous ...multiplexing) Requires large cell population, senstive to time since cell fixing HTGTS Not reported Cell-based...sequencing techniques, please see “Methods for Optimizing CRISPR-Cas9 Genome Editing Specificity” by Tycko...Vic E. Myer, and Patrick D. Hsu. "Methods for Optimizing CRISPR-Cas9 Genome Editing Specificity." Molecular...
  18. Antibodies 101: Validation

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    Blog Post
    ...inconclusive results, erroneous conclusions, and wasted time. In fact, many have called out antibodies as major...and so this approach may not be best if you are ultimately planning to use your antibody to detect endogenous... proceed with confidence.  Share your results Ultimately, like all of science, validation experiments ...could work great in another context! So the next time you test an antibody (perhaps one from Addgene!)...
  19. An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency

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    Blog Post
    ...with linear DNA as repair templates which reduces time to generate edits to only four days (Paix et al.... rapid. This stringent selection allowed me to optimize a range of editing parameters: oligo repair templates...Farboud et al., 2019). The Meyer lab also determined optimal PAM orientation for editing using two Cas9 targets...Resources Learn about gRNA design and gRNA optimization for your CRISPR experiment ...
Showing: 421 - 440 of 923 results