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Showing: 121 - 150 of 922 results

  1. Guide to Using Pooled Libraries

    Type
    Guide
    ...can help you begin to understand what genes are important to a certain phenotype, and allow you to design...the screening process. Amplify the library (electroporation and maxiprep). If delivering as virus, make...die or do not pass selection (in the case of a reporter) Sort “winning” cells Sequence those cells that...

  2. Promoters

    Type
    Guide
    ...silencers. Promoter Regions There are three main portions that make up a promoter: core promoter, proximal...promote the initiation of transcription. The most 3' portion (closest to the gene's start codon) of the core...transcription factors bind. Distal Promoter The final portion of the promoter region is called the distal promoter...

  3. Adenovirus Guide

    Type
    Guide
    ...first-generation rAdV vectors than later generations, which incorporate additional genomic deletions. Figure 2: First-generation...Adenovirus Vector as a Zika virus vaccine . Scientific Reports, 10 (1), 3613. https://doi.org/10.1038/s41598-020...adenoviral genes provided in trans . Seroprevalence Proportion of individuals within a population that present...

  4. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ...customized fluorescent reporters can be challenging enough, but incorporating multiple reporters into a single ... the 3’ UTR of the reporter mRNA via an MS2 coat protein (NLS-MS2-RFP) the reporter mRNA, which contains...their reporter mRNAs. Using the TRICK plasmids, Halstead et al demonstrated that 91% of reporter mRNAs... used to study transport characteristics of various Golgi and plasma membrane reporters as well as secretory... protein transport. Franck Perez’s lab has developed a whole collection of hook and reporter proteins ...pyralis) reporter plasmids for use in high-throughput screening of pre-mRNA splicing. This reporter system...widely used luciferase reporter gene plasmid pGL3 pGL3 luciferase reporter gene plasmids have been...

  5. Using Phosphoserine to Study Protein Phosphorylation

    Type
    Blog Post
    ...the pSer-incorporated polypeptide. These three components were shown to effectively incorporate phosphoserine...Creating phosphoserine-incorporated proteins The generation of pSer-incorporated proteins requires a molecular...strain for pSer-incorporation The second component necessary for phosphoserine incorporation is a modified...considerations, it is important to determine how much phosphorserine has actually been incorporated into the full-length...different levels of incorporation due to differences in stability of the p-Ser incorporated protein as well...determine the stoichiometry of phospho-incorporated to non-phospho-incorporated proteins purified from the system...pSer-incorporated to non-phosphorylated protein. Verification of site-specific pSer incorporation can ...

  6. Using AAV for Neuronal Tracing

    Type
    Blog Post
    ... facilitate transport: retrograde transport relies on dynein, while anterograde transport relies on kinesin...take advantage of existing transport pathways in neurons. Anterograde transport is typically used for the... transmitter synthesis. Retrograde transport is used for transport of endocytosed material or molecules... impaired, and only few reports exist about dye molecules that are transported from one neuron to another...There are several reports of both retrograde and anterograde intracellular transport of some AAV serotypes...including dendritic and axonal extensions and, most importantly, any connections the neurons make to other distant...for various forms of retrograde and anterograde transport suggest that there are several parallel mechanisms...

  7. Replacing Paper: Tips for Choosing an Electronic lab Notebook

    Type
    Blog Post
    ... whether respondents saw them as not important to very important. In addition, we also explored the barriers...”. The study also showed that It is important that an ELN supports all types of data, which enables researchers...electronic ones? It is important to know in which ways your data can be exported or printed from the ELN...project. Can you meet the ELN support team? We cannot stress enough how important this is when deploying an...only 46% could be reproduced. In a recent Nature report from researchers at Amgen, it was stated that only...through all saved data and automated writing of reports. To be able to understand the situation in depth... is likely to require on-going maintenance and support.” Besides that, academic institutions tend to think...

  8. Antibodies 101: The Four ELISAs and When to Use Them

    Type
    Blog Post
    ...conjugated with a reporter or tag that can then be detected.  Typically, the reporter is an enzyme such...indirect and sandwich ELISA the reporter signal is directly proportional to the level of antigen in the...whereas in a competitive ELISA the reporter signal is inversely proportional to the level of antigen in the...dish and incubate it with a reporter conjugated primary antibody. The reporter reacts with a provided substrate...simplicity we will discuss the more common enzyme-based reporter methods. For enzymatic signal detection, users... sandwich and competitive. In a direct ELISA a reporter enzyme conjugated antibody binds to antigen coated...antibody first binds to the antigen and then a reporter enzyme conjugated secondary antibody binds to ...

  9. The Many Reasons for Irreproducible Research- and a Vaccine to Eradicate It

    Type
    Blog Post
    ...you think there are four important times to measure that growth rate, you report the results of all four... registered reports, the gold standard for testing hypotheses Finally, Registered Reports combine all ...Reproducibility Project: Cancer Biology, where reported methods may not include how many tests were conducted...attention on the results instead of the methods. Unreported flexibility and questionable research practices...result after conducting a replication study. An important difference between the original setting and the...genetic variation, or different handling can all be important.  If we eliminate the problems above: unclear ...research finding we can be more certain that some important, previously unknown variable actually makes a ...

  10. Finding nucleic acids with SHERLOCK and DETECTR

    Type
    Blog Post
    ... to cleave and activate a reporter. The detectable signal from this reporter can be measured and quantified...and a Csm6 specific reporter (in the same fluorescent channel as the Cas13 reporter), they could significantly...variants, Cas13, and Cas12a, to develop simple, portable, and inexpensive platforms to reliably detect ...method termed Specific High Sensitivity Enzymatic Reporter UnLOCKING (SHERLOCK and SHERLOCKv2) (Gootenberg... termed DNA Endonuclease Targeted CRISPR Trans Reporter (DETECTR) (Chen et al., 2018). Both SHERLOCK and... SHERLOCK- Specific High Sensitivity Enzymatic Reporter UnLOCKING Cas13 (C2c2) was first identified in...molecules. In SHERLOCK a quenched fluorescent ssRNA reporter is added to the reaction. Cleavage of the quenchable...

  11. Visualizing Translation at the Single Molecule Level

    Type
    Blog Post
    ... the 3’ UTR of the reporter mRNA via an MS2 coat protein (NLS-MS2-RFP) the reporter mRNA, which contains...removal of GFP from the TRICK reporter. Delayed translation is known to be important in development, and Halstead...proteins to allow translation to occur. For a TRICK reporter mRNA this displacement leads to a change in the...GFP protein that binds to the coding region of a reporter mRNA via a PP7 coat protein (NLS-PCP-GFP) an RFP...translation, both GFP and RFP are bound to the reporter, causing it to appear yellow. The act of translation...of the mRNA, leaving only a red FP bound to the reporter. Because it contains a nuclear localization signal...copies of the FP-coat protein fusions to their reporter mRNAs. Using translation inhibitors cycloheximide...

  12. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    ...the promoter of the reporter gene to activate expression (Figure 2c). The reporter gene then allows detection...genes as reporters and some that repress instead of activate transcription. The HIS3 reporter system (...sequence (UAS) of its target gene, in this case a reporter gene (e.g. luciferase or GFP), regardless of whether...co-expressed in a yeast strain that also contains a reporter gene cloned downstream of a promoter containing...referred to as the “bait”, Figure 2b) cannot activate reporter gene expression on its own, but, if the two POI...Gal4 activator, bacterial LexA DBD and the lacZ reporter gene (3, 4), there are now many variations, including...to have Y2H systems that avoid transcriptional reporters altogether; In the split-luciferase system, intein-mediated...

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

    Type
    Blog Post
    ...cuts to important research funding.  So when I saw that there was going to be a march in support of science...the rain, about 4,000 supporters of science showed up to march. We had full support of local law enforcement...proved to be the perfect opportunity to cement in my children's heads the importance of science and positively...seems to break everyday. Many researchers and supporters of science marched to advocate for scientist ...for science, and numerous other reasons. It is important to remember that we, scientists and non-scientists...°W – Vanessa Sung Hundreds of scientists and supporters in Montreal took to the streets on April 22 to...Albany, Rep. Paul Tonko also came and spoke in support of science - he's one of the few members of Congress...

  14. Technologies Enabled by NanoLuc® Luciferase

    Type
    Blog Post
    ...has dubbed these fusions LumiFluor Reporters. The LumiFluor reporter constructs were designed to be packaged... The researchers took a FRET reporter and adapted it into a BRET reporter by using NanoLuc® Luciferase...share your constructs from lab to lab. Promega supports the Addgene mission statement: Accelerate research...(FLuc) and Renilla (RLuc) luciferase. This is important not so much for how bright you can make a reaction...couples NLuc to fluorescent proteins to make better reporters and the second couples NLuc to fluorescent proteins...host cells. The fluorescent moiety of the BRET reporter was used to sort successfully infected/expressing...NanoLuc luciferase. Both GpNLuc and OgNLuc BRET reporters produced bright signals without exogenous excitation...

  15. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    ... to report mitochondrial turnover Mitochondrial health, transport and turnover are important features...genes, reporters, or other markers into a target genome. They did this by selecting the important elements... be imported to the mitochondrial matrix. Gottlieb’s lab has used an inducible MitoTimer reporter using...usefulness in cell culture to report changes in mitochondrial turnover and transport. pMitoTimer from Yan’s ...s lab, which is constitutively expressed, reports on the balance of biogenesis and mitochondrial degradation...mitochondria in the networks marks the differences in import of newly synthesized (green) protein and organelles...channelrhodopsins. CnChR1, nicknamed Chrimson, is the first reported yellow-peaked channelrhodopsin with a spectral...

  16. Transferable Skills Guide: Managing a Team

    Type
    Blog Post
    ...mentor Your manager should support you, just like you are supporting your report(s). This means that you ...unique skills and strengths (you will likely have reports with different levels of experience), find out ...how to prioritize but I  just added the “most important” tasks my schedule. This was bad for 2 reasons...tasks before I was a manager! Second, my direct report was not engaged because I only left him/her with...expectations, answer any questions, and (most importantly) decide on a timeline. My experience is that ...you’re delegating a recurring task to a seasoned report, there should be questions! Remember that delegating...develop your team members by giving them new opportunities. I still need to check myself sometimes to ...

  17. Experimenting with New Careers while in Grad School

    Type
    Blog Post
    ...of the opportunities you’re provided throughout your PhD program.   Find people who will support your ...had a wonderfully supportive mentor and graduate office. I cannot stress how important they were to my ...experimenting with the career and networking opportunities available to me as a graduate student culminated...events for adults and professional development opportunities for researchers interested in connecting with...they had my back. If your advisors are not as supportive of your career goals, find people who are. I ...as types of experiments. I searched for new opportunities with an open mind and open ears. Everyone I ...mentioned, my graduate office staff was amazingly supportive. I talked to them about my interests and my ideas...

  18. Celebrating 15 Years of Scientific Sharing

    Type
    Blog Post
    ...everywhere can access plasmids despite the evolving export/import rules. Our shipping and lab teams continue to...entire supportive Board, we have a robust committee process in place and continue to incorporate additional...all of the success we have achieved and the opportunities we have for the future. At Addgene we will never... The success of these projects allows us to support more new projects. Diversification is happening... quality and success. Customer and technical support Our customer and technical service brands continue...sharing, but we don’t have to worry about getting support from depositors. We regularly hear how much they...blessed with a large community of scientists who support us by depositing, requesting and spreading the ...

  19. Networking and Inspiring Young Scientists at Freshers Fairs

    Type
    Blog Post
    ...benefit from some Addgene support? Please get in touch, as we aim to support a couple of events a year...across Europe get introduced to the cultures and opportunities available to them at university. At a Freshers... into university life. The fairs are also an opportunity for the societies to promote their upcoming events...biological societies that Addgene has had the opportunity to work with at Freshers Fairs in the past year...undergraduates and PhD students so that they can help support each other and provide advice. The society organizes...involved with. The society provides talks and career support for a range of different scientific disciplines... in its first year. The society facilitates opportunities to learn about Synthetic Biology and exciting...

  20. 27 Hot Plasmids from 2016

    Type
    Blog Post
    ...et al. Cell. 2015. PubMed PMID: 26627737 pORTMAGE: A portable method for bacterial genome engineering ...-ome using a quick, affordable, and ubiquitous reporter assay. In their recent publication, Kroeze et ...their enhanced TANGO arrestin recruitment assay incorporating 314 codon-optimized GPCR sequences for human...MAGE uses recombineering (2) to simultaneously incorporate multiple single-strand DNA (ssDNA) oligonucleotides...so-called “genomically recoded organism” (3) but its portability to other bacterial strains remains seriously ...bacterial species (4). This set of plasmids (dubbed pORTMAGE) expresses the λ Red recombinase enzymes, as well...can be used in a broad range of strains. Thus, pORTMAGE simultaneously allows genome editing and mutant...

  21. Expanded Access to Rabies Plasmids

    Type
    Blog Post
    ...plasmids, are export-controlled materials in the USA, where Addgene is located.  The export license, if ...requestors. We also offer export license applications for VSV and SARS-COV-1 plasmids. Export control license ...Addgene is excited to announce we now offer an export license application for rabies materials! Some ...license process    An export control license, once approved, allows Addgene to ship plasmids or viral vectors...the applicable research project when purchasing export-controlled materials. Addgene will then apply for...process.   Due to recent regulation changes, an export control license for rabies materials may be required...required for others. You will be notified if an export license is not required for your country after ...

  22. Advice on Career Paths and the Green Card Process for International Researchers and Entrepreneurs

    Type
    Blog Post
    ...You need the support of your institute to do this one. Universities will usually only support tenure track...Further Evidence (RFE). Letters of support are the only opportunity that you are given to present your...and so on. Candidates should look broadly at opportunities to develop skills related to their areas of ...must have both substantial merit and national importance; (2) The foreign national must be well positioned...evaluating an NIW, USCIS will also focus on the importance of the individual’s work in their field and the...working under in the lab or has won, awards, media reports, and membership in groups related to the foreign...This criterion is best proven through letters of support from experts in the field. You must also be able...

  23. Fluorescent Proteins 101: Luciferase

    Type
    Blog Post
    ...Fluorecent Proteins in Nano Lanterns Reporter gene assays A reporter gene, such as luciferase, usually serves... is also the most commonly used bioluminescent reporter. This beetle emits a yellow-green light with a...several studies utilized luciferase as a genetic reporter in plant and mammalian cells. Luciferase assays...available to choose from) is now a common feature in reporter plasmids.  Learn How Luciferase Can Be Used In...transcription within cells, where detection of the reporter protein or its enzymatic activity is measured....expression can be determined by detection of the reporter in a specific assay, which ideally would have .... Production of photons by this bioluminescent reporter occurs slower than fluorescent-based methods, ...

  24. 7 Tips to Secure a STEAM Internship This Summer

    Type
    Blog Post
    ...STEAMid. Internships open doors to a web of opportunities that the classroom cannot offer. Furthermore... not deter any student from applying to the opportunities of their dream.  To help students focus on the...below:  1. Eligibility check Scrutinize every opportunity prior to starting an application to assess your...unless you are applying for a Federal Government opportunity, which tends to be more detailed. Beyond the ...draft a personal statement, which is the perfect opportunity to paint a comprehensive picture of your academic...statement will pay dividends. This is the only opportunity where you get to impress the judges on your ability...your recommenders deliver a superb and detailed portrayal of yourself. Be mindful that January and February...

  25. Quick Guide to Near-Infrared Fluorescent Proteins

    Type
    Blog Post
    ...molecular brightness (Table 1).  NIR reporters and biosensors NIR reporters can tell us about cellular events..., D. & Michnick, S.W. An infrared reporter to detect spatiotemporal dynamics of protein-protein interactions...FPs) were derived from natural photoreceptors incorporating biliverdin IXα (BV) autocatalytically. Fortunately...should be supplied (Baloban et al., 2017).  It is important to consider effective brightness of NIR FPs in... a cell and depends on the efficiency of BV incorporation by an apoprotein and protein expression level...higher apparent cellular brightness than previously reported widely used dimeric iRFPs (Table 1). In general...Oliinyk et al., 2019). They were engineered to incorporate BV. Their advantages are their small size (17...

  26. 3 Tips to Improve HDR Efficiency for CRISPR Editing in Human Cells

    Type
    Blog Post
    ...it is incorporated. We were surprised to see how rapidly the efficiency of mutation incorporation falls...contains the intended sequence changes to be incorporated into the edited genome. In mammalian cells such...these two strategies allows you to specifically incorporate homo- or heterozygous mutations. 1. Increasing...therefore strongly encourage any researcher to incorporate CRISPR/Cas-blocking mutations at both targeted... our frustration) in many cases we only saw incorporation of the blocking mutation. Despite the fact that... away from the cut site were not efficiently incorporated during HDR. We decided to systematically characterize...only about 30 bp, it is no longer feasible to incorporate mutations without screening thousands of clones...

  27. Hot Plasmids April 2018 - Protein Degradation, Nanoscopy, FIRE-Cas9, and Yeast Expression Tools

    Type
    Blog Post
    ...expression, sometimes incorporating chemically inducible approaches to provide temporal control, this new ...random variation, or noise, in reporter expression. To do so, one places reporter expression under the control...expression of a reporter protein, it is possible to change the amount of noise in reporter gene expression...can deliver these components together using electroporation. In these bulk cell experiments, Clift et al...no stress or damage upon TRIM-Away, and the electroporated cells re-adhered to the culture plate within...deliver the target antibody. TRIM-Away reagents and reporters are available from Addgene - we hope they will...lab has deposited plasmids encoding fluorescent reporters and other proteins controlled by GAL4 and ZIF268...

  28. Gendered Innovations: Why Does Sex of the Cell Matter?

    Type
    Blog Post
    ...emphasizes that reporting the sex of cells and tissues—when that information is reliable—is important to ensuring...Universities need to incorporate these methods into their curricula. Corporations need to integrate these...Governments, universities, and, increasingly, corporations in the US and Western Europe have taken three...into research. Why are sex and gender analysis important? Doing research wrong costs lives and money. According...positive, the analysis did find 263,000 more osteoporotic fractures. The overall economic savings to the... goal of gendered innovations It is crucially important to get the research right from the beginning. ...unique to male stem cells. Nor did they detect important differences in function between male and female...

  29. Plasmids 101: FLEx Vectors

    Type
    Blog Post
    ...gene. Spatio-temporal expression of optogenetic reporters: Since most optogenetic reporters are delivered...invert the reporter and GeneR fragment. Then the second recombination event would excise the reporter from ...transduced reporter gene. Generating a FLEx switch to control expression of an optogenetic reporter would ...Cre-dependent reporter expression, the potential for background expression of the reporter will be greatly...shown in Figure 2, the cassette should contain a reporter DNA sequence (such as GFP) in the sense orientation...the two pairs of target sites. By flanking the reporter with one loxP site and one lox511 site in the ...would ensure that the reporter remains silent until a cell or tissue-specific Cre is provided. Since the FLEx...

  30. How to Design Your gRNA for CRISPR Genome Editing

    Type
    Blog Post
    ...sequence are important considerations for designing your gRNAs. For indels, it's not so important what location...nom. Me love gRNAs! Comic by Maya Kostman.   Important considerations before you start an experiment ...tool among them. While this seems obvious, it is important to remember that the same is true when designing...location in the gene you target, but it is important that your gRNA sequence is designed to be highly active...CRISPRi, these considerations are of roughly equal importance (target should be near the TSS but you can worry...from). Finally, for HDR, location is much more important because you have to target within ~30 nt of your...gRNA with an optimized sequence is of primary importance (more on this below). Created with BioRender.com...
Showing: 121 - 150 of 922 results