Skip to main content
Addgene

Enter a sequence to perform a BLAST-based search of Addgene plasmid sequence databases.

In Category

We narrowed to 991 results for: igh

Showing: 261 - 280 of 991 results

  1. Antibodies 101: Affinity Reagents

    Type
    Blog Post
    ... standard antibody: An ~150kD protein with both light and heavy chains made by immune systems. But here...imaging applications, smaller affinity reagents means higher resolution. Many of the below are also small enough...the lab, instead of generated in vivo, they have higher reproducibility and consistency in the final product...cleavage. They are comprised of the variable heavy and light chain regions, connected by a short linker peptide... longevity compared to antibodies, along with a higher likelihood of aggregation (Bates, 2019). They are..., are derived from a camelid antibody lacking a light chain, called a heavy chain antibody (Muyldermans.... DARPins typically consist of 3-4 repeats, have high specificity and binding affinity, can be produced...

  2. 7 Simple Strategies to Resolve Conflicts with Difficult Supervisors

    Type
    Blog Post
    ...supervisor might expect weekly or biweekly progress reports. However, a hands-off busy supervisor might get ...Know-it all experts, who think their way is the only right way, and Indecisives, who keep changing their ... your discussion will get side-tracked, and you might create even more conflicts. 4) Define in advance...about mutually beneficial solutions Your supervisor might not agree with all of your ideas, but he or she ... conflict is miscommunication. For example, you might misunderstand your supervisor’s suggestions and ...sense their tone of voice in an email, there is a high likelihood of miscommunication, which can lead to...assertive to get your point across. Some supervisors might try to intimidate you with negative comments (hostile-aggressives...

  3. RNA Interference in Plant Biology: New Tools for an Old Favorite

    Type
    Blog Post
    ...technologies, one might incorrectly relegate RNAi to history. The straightforwardness of RNAi is unmatched—unlike...considered the “gold-standard” for LOF studies, the high gene copy number found in plants makes traditional... DNA, thereby limiting CRISPR’s effectiveness at high copy numbers and genomic loci with low accessibility...directly compared CRISPR to RNAi in the context of a high throughput survival screen found that either technology...for cell growth. Together, this excellent work highlights the importance of using orthogonal methods to...Arguably the greatest advantage of RNAi is the straightforward nature of the technique—all that is required... long hpRNA with Gateway-based cloning and straightforward positive selection of plants undergoing RNAi...

  4. Sequencing Options for CRISPR Genotyping

    Type
    Blog Post
    ...information from targeted amplicon sequencing is highly sensitive with detection levels as low as .01% ...of <4 bp. The off-target list is also generally weighted by the position of the mismatch along the length...detect double-stranded breaks and do not require high sequencing read counts making them fast and viable...Searches the genome for all DSBs, doesn't require high read counts, fast multiplexing Requires delivery...cells Not as senstive as other unbiased methods, high background BLESS Not reported Cell-based (in vitro...component required (e.g. dsODN), doesn't require high read counts (fast multiplexing) Requires large cell...can evaluate and validate off-target sites in a highly accurate manner. Using both of these approaches...

  5. Production of Virus in Insect Versus Mammalian Cells

    Type
    Blog Post
    ...provided some insight on these differences in viral production and will help you decide which is right for you...available, or the cost of custom viruses are too high for your budget. In those cases, you’ll need to ...more common to make lentiviruses and AAV in-house. High-titer adenoviruses are difficult and time consuming...be produced in a greater volume and will have a higher titer than a P1 virus. Pro-tip! Centrifuge the ...this is only possible because baculoviruses are slightly less prone to recombination than other viruses...Processing Processing of your baculovirus is straightforward — simply spin down the supernatant in a centrifuge...the next viral passage, titer the virus, or go straight into your viral application, like protein production...

  6. Academic vs. Industry Postdocs

    Type
    Blog Post
    ...in a workable direction quickly. Of course, you might have less time to explore scientifically since you...industry, which means you’re more likely to have the right contacts and get your resume pulled to the top of... job applications require teaching experience or high numbers of publications, which may take a longer...chances of getting a good manager do tend to be higher in an industry setting. Scientific community and...and mentorship: As an industry postdoc, you might find yourself feeling scientifically isolated if no... have an in-depth discussion about your project might be challenging. In an academic environment, surrounded...re more likely to give an oral presentation to highlight any new data or ask for additional resources, ...

  7. CRISPR 101: Mammalian Expression Systems and Delivery Methods

    Type
    Blog Post
    ...protein Nucleic acids Nucleic acids Throughput High High High Efficiency Medium Medium Medium Difficulty...acids or protein Throughput High Very low Efficiency High Very high Difficulty Very easy Very hard...product in mind, steps 1 and 2 will generally be straightforward. Likewise for step 4, as this ties directly...a mammalian cell line that can be transfected at high efficiency. Lentiviral transduction Cas9 and gRNA...been published as CRISPR testing tools: Traffic Light Reporter System: Can evaluate lentiviral component...

  8. Addgene's Top Blog Posts from 2020

    Type
    Blog Post
    ..., we've looked back on the year would like to highlight some of the top articles from the Addgene blog...-CoV-2 protein expression, and more. We also highlighted reagent repositories and other collaborative ...time so that you can begin your cloning experiment right away. And without further ado, here are the top ...about hosting multiple plasmids in one cell, it might be that you can't stably maintain them in the cell...free molecular biology tools out there! Here, we highlight a subset of these tools for plasmid mapping, DNA...

  9. Why Do I Need an MTA Anyway?

    Type
    Blog Post
    ...purposes Rights to all research results, modifications, and inventions created by the Recipient Right to ...terms, so you can rest easy knowing your rights – and the rights of your institution – are protected while...scientist working hard on research, and you need the right experimental materials. Who has time for paperwork...materials. Questions about intellectual property rights, pending publications, and commercial use often...patent applications on modifications or inventions Right to publish without editorial comment or review by...order, Addgene generates the MTA and emails it straight to you, or we can send it directly to your institution...

  10. What’s in Your Tube?: A Quick Guide to Measuring DNA by Spectrophotometry

    Type
    Blog Post
    ...but some details might vary.  How it works — briefly Spectrophotometers measure how light travels through...through a sample — in this case, the absorbance of light at three key wavelengths: 230 nm, 260 nm, and 280...comparison. Each of these contaminants will have a slightly different impact on the spectra of a mixed sample...molecules that may be present in the sample also absorb light in this range of wavelengths (Figure 1B-D). These...remove from your sample is protein. Proteins absorb light to varying degrees across the UV range used in NanoDrop...This property means that even samples with quite high levels of phenol (up to 600 ppm according to Loughrey...difficult by spectrophotometry. RNA tends to yield a higher A260/280 and a lower A260/230 than DNA, so shifts...

  11. With an Eye Towards the Future, We Look Back at the March for Science

    Type
    Blog Post
    ...marched in solidarity with our American neighbours, the fight is far from over here at home.  The work...of which impact reproductive rights and justice. Whether it’s used to fight the myths that abortions cause...wishful thinking. The March for Science is about fighting scientific partisanship, including more scientists...there's been a disturbing trend in the number of high profile people denying science, from celebrities...convince me to spend a Saturday in the tourist-trap nightmare that is Times Square. But I made the sacrifice...Most people won't argue with that idea, but they might feel disconnected from the entire scientific process...I think many of us grew to assume that the data might speak for itself but we're now in a rift where our...

  12. Fluorescent Proteins 101: Aptamer Fluorophores

    Type
    Blog Post
    ...RNA molecules, which could bind their targets with high specificity and affinity, were eventually called...nucleotide-based ligand binders would prove to be highly adaptable tools. The key to aptamers’ flexibility...conformational changes after target binding, aptamers are highly customizable in terms of how they actually “report...applications. Learn About Fluorescent Biosensors Light-up aptamers (or, fluorescing with veggies) Though...eventually evolved into Spinach2, a more stable and brighter version of the aptamer, but it still had limitations...more stable in cells. The new aptamer also has a higher melting temperature than its Spinach ancestors,...have also made aptamers an important component of high throughput assays with microarrays and biochips....

  13. "Hall of Fame" AAV Enhancers from the Allen Institute for Brain Science

    Type
    Blog Post
    ...collection of AAV enhancer plasmids with high specificity and brightness, now available at Addgene!   {% module_block...winners, containing enhancers with the highest specificity and brightness. You will find enhancers to target...allocated tremendous time and resources to generate highly specific tools to target different cell populations...border-style":"none","padding-left":"172px","padding-right":"172px"}{% endraw %}{% end_module_attribute %}{...font-family":"helvetica","font-style":"normal","font-weight":"normal","text-decoration":"none"},"fallback":...font-family: helvetica; font-style: normal; font-weight: normal; text-decoration: none","font_set":"DEFAULT...best-in-class collection. Hopefully, this collection of highly specific AAV enhancers can help scientists around...

  14. Healthcare Consulting: A Door to the Business of Life Sciences

    Type
    Blog Post
    ... client situations might be especially challenging. For instance, the client might need to get a large...what a career in this field involves, and why you might enjoy doing it. I’ll also talk about the skills ...and anticipated commercial success. Or the client might need help with the intricate task of designing a...topics are typically cutting edge; a consultant might outline the impacts of combining targeted molecular...turnover of projects, pressure to maintain consistently high levels of performance, and occasionally being faced...cases to better staff future teams. As with any highly collaborative work place, challenges can arise ...or medical device works. This understanding sits right at the core of the suite of benefits a given therapeutic...

  15. Chemogenetics vs. Optogenetics: Which Method Should I Choose?

    Type
    Blog Post
    ...activation (chemogenetics) or light (optogenetics).   Timing Optogenetics offers a high degree of temporal resolution...presence or absence of the correct wavelength of light. This response is immediate, on the scale of milliseconds...tissue-subregion targeting down to the limits of light delivery. Chemogenetic approaches excel at broad...offers better control over these factors as the light source can be easily and quickly manipulated or ...Addgene website. Optogenetics also requires that light of the correct wavelength reach receptors in the... address this is by working with organisms with light accessible brains, such as zebrafish larvae. Mouse...implant to be surgically installed to allow for light delivery. Chemogenetics poses the advantage of being...

  16. An Addgene Summer Internship

    Type
    Blog Post
    ... the high school interns as equals, because as soon as I had passed my training, I dove straight into ... never would have imagined that my summer before high school senior year would have been spent within ...weeks that we had made a mistake by not having the right concentration of a reagent during our HiFi assembly...the following two weeks, we were able to obtain eight clones. We ran digested and undigested samples of...one clone we found with the correct bands was the right plasmid, so after discussion, we sent it for sequencing...invited guests came to see the research from all the high school interns, done within the past six weeks. ...other way. Toj Toj-Villagran is a rising senior in high school who just completed their summer internship...

  17. Hot Plasmids and Viral Preps - July 2021

    Type
    Blog Post
    ...fluorescent biosensor has a high sensitivity (50 nM-500nM range) and a high specificity for SLs in vitro...Every few months we highlight a subset of the new plasmids and viral preps in the repository through ...organoids. The Organoid EasyTag System is a straightforward workflow for doing gene targeting, reporter.... This novel type of biosensors could be used in high throughput screening for agrochemical compounds ...CRISPR Plasmids and Resources page. Here are some highlights from the past couple months: A new lentiviral...

  18. FastCloning: A Newer, Simpler Cloning Technique

    Type
    Blog Post
    ... tips for high efficiency cloning with this technique: Amplification efficiency is highest when using...labs, a variety of other cloning techniques with higher efficiency and simpler procedures have been developed... in the figure from Chaokun et al., 2011 to the right. After PCR, you should confirm that the products...introduced during PCR or any subsequent steps. Using high fidelity DNA polymerases, I have never had any mutations...chance of introducing an unwanted mutation. Tips for high efficiency cloning Now that you have a better understanding...using high fidelity DNA polymerases like phusion. An 18-cycle PCR leads to the most colonies after transformation...27391134. 2. Li, Chaokun, et al. "FastCloning: a highly simplified, purification-free, sequence-and ligation-independent...

  19. Using AAV for Neuronal Tracing

    Type
    Blog Post
    ...neurons in the mouse brain. MouseLight images can be found on the MouseLight website. Together with scientists...neuroanatomy and neural connectivity, providing valuable insights for neuronal communication. Viral vectors and...vectors can be equipped with strong promoters for high and neuron-specific transgene expression. Fluorescent...for their production and purification that yield high titers (see also AAV protocols for production, purification...researchers from the Janelia Campus for their MouseLight project but neurons were traced with AAV2. Two-Photon...variant, rAAV2-retro, that migrates retrogradely at higher rates than any other AAV serotype (Tervo et al....also having the added advantage of lower toxicity, higher specificity and reduced technical variability compared...

  20. Thoughts on the Future of Research 2015 Boston Symposium

    Type
    Blog Post
    ...forward, in this post I’ll point out some of the highlights and takeaways I got from the symposium. The ...outcome is not easily achieved. Of note, it was highlighted during the conference that just increasing funding...projects sought to make an interactive game that highlights just this point. As part of this, trainees need...thoughts The FOR Symposium Boston 2015 shined a light on some of the many problems facing the academic...academic research enterprise. In addition to highlighting some of the important questions that still need to...
Showing: 261 - 280 of 991 results