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We narrowed to 1,052 results for: FER

Showing: 281 - 300 of 1052 results

  1. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    ... the split-luciferase system, intein-mediated splicing reconstitutes functional luciferase only when fusions...when separated (2), and that DBD and TAs from different systems could be combined effectively (3). In ...target gene, in this case a reporter gene (e.g. luciferase or GFP), regardless of whether it interacts with...sequence of the DBD. The DBD-POI fusion (commonly referred to as the “bait”, Figure 2b) cannot activate reporter...interact, this will bring the POI-TA fusion (commonly referred to as the “prey”) in close enough proximity to... fusions of the two halves of the luciferase protein interact and enable intein splicing (5). Screening...a more vigourous screen is to use a number of different bait and prey vectors. This has been shown to ...

  2. Promoters

    Type
    Guide
    ... Plasmid Reference Molecular Biology Reference Promoters Promoters Definition A promoter is...Addgene's molecular biology reference material covering promoters, transcription and the transcription...mRNA transcript produced. The antisense strand is referred to as the template strand or non-coding strand...transcription of DNA into mRNA. Promoter binding is very different in bacteria compared to eukaryotes. In bacteria...promoter region. Each sigma factor recognizes different core promoter sequences. Operons Although bacterial...regulation, like operons. Operons are a cluster of different genes that are controlled by a single promoter... types of RNA polymerases that all transcribe different genes. RNA polymerase I transcribes genes encoding...

  3. Viral Vectors 101: Pseudotyping

    Type
    Blog Post
    ...virus:  Lowering cytotoxicity. Different envelope proteins have different levels of cytotoxicity and the...Altering sensitivity to serum. Different pseudotyped viruses are differentially inactivated by nonspecific...Table 1 (Cronin et al., 2005)     References and Resources References: Crawford KHD, Eguia R, Dingens AS...protocol for generating lentivirus, but you use a different envelope glycoprotein instead of the wild-type...Lentiviral production uses three plasmids: (1) The transfer plasmid, (2) the envelope plasmid, and (3) the...review articles to find tables and information on different viral membranes, tropism, and advantages and disadvantages... Jennifer Tsang...

  4. Bacterial Toxin-antitoxin Systems as Molecular Biology Tools

    Type
    Blog Post
    ...more stable than the methyltransferase. When a plasmid is lost, the methyltransferase is degraded and/or...of CcdB (toxin) and CcdA (antitoxin) where CcdB inferes with DNA replication by binding the GyrA subunit...composed of restriction endonucleases and methyltransferases. Restriction endonucleases cleave DNA at ...specific nucleotide sequences. To counter this, methyltransferases add methyl groups to bases at specific nucleotide...address a diverse set of biological inquiries. References Ogura, Teru, and Sota Hiraga. "Mini-F plasmid...genes that couple host cell division to plasmid proliferation." Proceedings of the National Academy of Sciences...Cara E., and James B. Kaper. "Use of stabilized luciferase-expressing plasmids to examine in vivo-induced...

  5. Tips and Tricks for Using Golden Gate Modular Cloning (MoClo)

    Type
    Blog Post
    ...experiments. The term ‘insert’ refers to plasmids carrying each part, ‘vector’ refers to acceptor plasmids, and...MoClo guide for all the basics of the process. Also refer to Figure 1 for a visual overview! Figure...vectors and parts you will be using during the different levels of your MoClo assembly. Know your process...dictated by four-base overhangs that are paired with different part types (e.g. 5’-GGAG and 3’-TACT flanking ...ensure that these would be interchangeable between different laboratories. Therefore, choosing acceptor vectors... Example Level 1 assembly. The choice between different Level 0 vectors depends on the four-base overhangs...MoClo plate, always do so in cold conditions and preferably on dry ice. Careful use of your glycerol stocks...

  6. Plasmids 101: Shuttle Vectors

    Type
    Blog Post
    ...protects against any of them. But different aminoglycosides work on different host species: kanamycin targets... unrelated host species, even from completely different kingdoms of life. That’s convenient for researchers...experiments in complex eukaryotic cells, but would prefer to grow, clone, and manipulate their vectors in...with BioRender.com. How are shuttle vectors different from other plasmids? If we all use the same molecular... Shuttle vectors have to address several big differences in the ways vectors are used across species: ...resistance gene on your vector might require two different selection molecules! It all depends on how the...Auxotrophic selection Another option is to use a different method of selection entirely. Auxotrophic selection...

  7. Twenty Years of Sharing: Addgene's Viral Vector Service

    Type
    Blog Post
    ... of the vectors in Addgene’s catalog are offered in different serotypes. In fact, the 879 viral vectors...these tools can be to use. AAVs, which can target different cell types depending on their serotype, are particularly...to researchers around the world. While we still offer many resources for producing vectors from plasmids...93,491 viral vector preps with researchers in 45 different countries — and as of 2024, we’ve distributed ... CRISPR, where we dove fairly deeply into the different types of tools scientists were using/depositing...prioritization happens for good reasons. We began offering our viral vector service because we realized that...successive generations developed ever since. GCaMP6 was offered through our viral vector service in 2016, and it...

  8. Viral Vectors 101: Parts of the AAV Packaging Plasmid

    Type
    Blog Post
    ...gene which differ between the AAV serotypes are responsible for serotype-specific differences in tissue...blog post to learn more about the AAV transfer plasmid! References: Naso MF, Tomkowicz B, Perry WL III...proteins needed for the virus to replicate,  and the transfer plasmid which contains the viral genome. In today... proteins. Alternative splicing generates the different C-terminal regions of the Rep proteins. While ...Replication initiates from the AAV genome ITRs on the transfer plasmid, which contain regions of dsDNA. To generate...the AAV p40 promoter. Alternative splicing and differing translational start sites generate the three VP...was engineered Learn about the parts of the AAV transfer plasmid  Resources on Addgene.org Learn about...

  9. Using Ultrasound to Image Bacteria in vivo: Acoustic Reporter Genes

    Type
    Blog Post
    ...Administration of Substrate: Some luciferase reporters, such as Renilla luciferase (RLuc) or NanoLuc, require...bacterial reporters that use fluorophores and luciferases to track bacteria in real time, but each of these...together genes from gas vesicle operons from two different bacteria, Bourdeau et al created two constructs... 1 (arg1) and acoustic gene 2 (arg2). The key difference between arg1 and arg2 vesicles is the amount ... invisible to ultrasound, which allows for differentiation of arg1 and arg2 ultrasound signals. This also...and arg2 plasmids are available from Addgene! References 1 Bourdeau, R.W., Lee-Gosselin, A., Lakshmanan...Blog Learn more about in vivo imaging tools The luciferase reporter gene The green fluorescent protein reporter...

  10. Antibody Neutralization Response Against Pseudoviruses Expressing SARS-CoV-2 Spike Protein Variants

    Type
    Blog Post
    ...19 pandemic,” says Balazs.     References and Resources References Cele S, Gazy I, Jackson L, Hwa S-...neutralization effectiveness of current vaccines differed depending on the variants (Garcia-Beltran et al...plasmids which encode spikes found on all of the different circulating variants. This allowed us to create.... In their assay, the pseudoviruses express a luciferase reporter which is used as a readout for infection...variants. Yet, the B.1.351 variant, which has only 9 different mutations compared to the wild-type, escaped neutralization...Garcia-Beltran et al., 2021 with permission.   Difference in neutralization after one dose and two doses..., this study was the first to compare so many different strains. It was also the first to perform neutralization...

  11. Single-cell tracking of lineage and identity with CellTag

    Type
    Blog Post
    ...Direct reprogramming describes the process where differentiated cells are turned into a cell type of choice...identifier and allows cells to be tagged with different CellTag libraries at multiple timepoints during... (iEPs), a self-renewing cell type that can differentiate into hepatic or intestinal cell types. MEFs ...inefficient. Cells were sequentially tagged with different CellTag libraries at three time points during ...MettI7a1 encodes an uncharacterized putative methyltransferase. Adding Mettl7a1 to the standard Foxa1-Hnf4α...are undergoing identity changes, such as the differentiation of induced pluripotent stem cells or the clonal...visualization tool. Check out the CellTag webpage! References Biddy, Brent A., et al. "Single-cell mapping ...

  12. K. phaffii: Rising to the Occasion

    Type
    Blog Post
    ...have risen into yeast expertise! References and Resources References Liu, Wei, et al., From Saccharomyces...applications in food science and biotech due to its fermentation abilities and other useful traits. The “biotech...lists some of the biological commonalities and differences between K. phaffii and other yeast strains.   ...aerobe Number of chromosomes 16 3 4 Preferred carbon source Glucose Glucose Glucose, methanol...Applications for K. phaffii There’s a number of different uses for the phaffii-lous strain. Protein expression...peroxisome studies since their peroxisomes will proliferate upon a media change into methanol and peroxisomal...peroxisomal studies on biogenesis, degradation, and proliferation. Genome manipulation K. phaffii’s genome has...

  13. Plasmids 101: How to Verify Your Plasmid Using a Restriction Digest Analysis

    Type
    Blog Post
    ...add loading buffer to your digest reactions before loading them. The glycerol in the buffer will make sure...based on the predicted sizes and organization of different features within the plasmid. Restriction analysis...can search NCBI for YGOI to find the particular reference sequence if necessary. The example plasmid on ...fragment having increased intensity to serve as a reference band. The uncut DNA (lane 2) shows 3 possible ...visualize the full size of your construct. Consider buffer and temperature compatibility when digesting with...versions of the endonucleases and/or supply custom buffers as means to avoid this issue. For your gel: ...in the gel well and the dyes provide a visual reference point so you can easily assess how far the gel...

  14. Plasmids 101: Screens vs. Selections

    Type
    Blog Post
    ...in. The differences They say the devil is in the details - and that’s where the differences in these... RNAi resources and go fishing! References and resources References Wang, T., Wei, J.J., Sabatini, D....? If you’re thinking, ‘wait, those terms mean different things?!’, you aren’t alone! We’ve noticed a lot...selections The common goals The confusion in differentiating between a screen and a selection isn’t surprising... you would individually treat cells with many different drugs and then perform an assay to look at an ...enhancer/suppressor screens. Selections Selections differ from screens in that only survivable manipulations...experimental question. Now that you understand the difference between the two, it’s time to check out our CRISPR...

  15. Antibodies 101: Monoclonal Antibodies

    Type
    Blog Post
    ...antibodies differ from other types of antibodies? How monoclonal antibodies are made “Monoclonal” refers to ...for your experimental needs.     References and resources References Bradbury ARM, Trinklein ND, Thie ...experiments, you may be curious about all of the different kinds of antibodies that are available. One common...heterogenous mixture of antibodies that recognize many different epitopes on the same protein - are more likely...to produce monoclonal antibodies. Genetic drift refers to changes in the nucleic acid sequence of the ...impacts the specificity, avidity, and number of different antibodies produced by the hybridoma. Image from..., Aldred SF, Bestagno M, Burrone O, Maynard J, Ferrara F, Trimmer JS, Görnemann J, Glanville J, Wolf P...

  16. Viral Vectors 101: Biosensors

    Type
    Blog Post
    ...imaging for advanced monitoring of different factors, in different or the same cellular location, all ...covered with packaging plasmids! References and Resources References  Bhalla N., Jolly P., et al. Introduction...biosensors provide quantifiable readouts for levels of different chemicals, hormones, proteins, etc., that you ...as calcium. This varied component, which we will refer to as the analyte, needs to be specifically recognized...and zinc. These ion sensors can be targeted to different cellular locations or tissues depending on the...acceptor of a FRET pair on actinin will produce different signals based on changes to the structure of actinin...organisms, but AAVs make it possible. In AAV systems, different serotypes can be used to affect or determine viral...

  17. From Lab Bench to Leadership: Transitioning into Management as a PhD

    Type
    Blog Post
    ...skills that were quite different from mine. Learning to bridge those differences was a pivotal moment in...technical problems in the lab, management involves a different skill set: mentoring, setting strategic goals,... day-to-day details. Ignoring individual differencesManaging a team isn’t “one size fits all”. Each team... needs. For instance, I had one colleague who preferred detailed instructions and regular check-ins, which...autonomously. Failing to recognize and adapt to these differences could have led to frustration, micromanagement... particular, served as a great "baby project," offering a supportive environment to build my management...noticed tension between two team members with differing approaches—one detail-focused and the other big-picture...

  18. Viral Vectors 101: Optogenetic Tools

    Type
    Blog Post
    ...excitatory and inhibitory channels activated at different wavelengths of light (Vierock et al., 2021). Step-function...stay on until a second pulse of light, with a different wavelength, turns it off, allowing the cell to... secondary signaling pathways. There are many different opsin (natural and engineered proteins responsive...pathway to activate various protein regulators at different points in the pathway. Photoswitches are great...successive steps along a pathway, there can be very different responses to long vs. short stimulation times ... & Cui, 2015). You’ll need to test for these differences when designing your experiment.  Finding the ...available to help light your path! Resources and references Recommended reading Abreu, N., & Levitz, J. (...

  19. Finding Your Perfect Job After University

    Type
    Blog Post
    ...’s of working in different sectors. Luckily, I have worked for a variety of different organizations, so...overview of my experience and how I felt about the different positions to enable them to decide for themselves...effectively. I had to learn how to cope with a different climate and culture, and also needed to be resourceful...very quickly; as soon as I learned how to do the different tests and became efficient, I then started to ...diagnostic tool for the detection and diagnosis of different types of pancreatic cancer and highlighting which...were keen for me to become involved with many different aspects of the company and gave me the freedom... and travel to meet scientists, making my job different every day. I love the freedom to design and develop...

  20. Building Global Connections with the International Mentorship Program USA-EUROPE

    Type
    Blog Post
    ...international mentoring conference held at the University of Valencia (Spain). At this conference, mentors and mentees...professional development and networking resources offered by programs like IMP USA-Europe not only provide...international mentors. Universities from five different geographical locations in Spain were enrolled:...mentees held scheduled monthly progress video conferences and had real-time correspondence via e-mail throughout...Mentorship Program and the International Mentorship Conference In addition to a pre-program self-assessment ... exchanged impressions of the IMP experience. Conference highlights included remarks by the Spanish Secretary...community overseas. Official representatives from different Spanish universities and high level public and...
Showing: 281 - 300 of 1052 results