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Showing: 541 - 560 of 1082 results
  1. Multiplexed Overexpression of Regulatory Factors (MORF) collection

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    Blog Post
    ...differentiation To understand how each human TF can contribute to differentiation, the Zhang lab transduced...Similar to the Human TFome Library, the MORF library can be used with other selection and screening mechanisms...library webpage. Disease models Good disease models can often be difficult to generate, in part because of... dysregulation. A targeted TF screening approach can not only lead to a better understanding of disease...simulate a disease in a model system. TF screens can be designed to identify custom TF programs to acutely...and GFP fluorescent controls. Alternatively, you can order individual plasmids from the library to further...Individual cellular fates were then matched with candidate TFs responsible for that differentiation.   ...
  2. Crowdfight, a Platform to Boost Scientific Collaboration During COVID-19 and Beyond

    Type
    Blog Post
    ...protocol can save weeks of trial-and-error. A recommendation for the correct statistical test can save a...Spain and works in the field of genetics and cancer. You can follow her on Twitter at @mahesa2504       ...community where both brief and long-term collaborations can be easily started and easily credited. And nothing...scientific research. It’s completely free, of course. You can find more information about us and what we do in ... based in Spain and works on climate change. You can follow him on Twitter at @IgnacioAmigoH     María...while most of the world was locked down, a South African company wanted to test if a type of antibodies ...
  3. Scientists Map the SARS-CoV-2-Human Interaction Network

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    Blog Post
    ... been motivated to do anything and everything we can to help find a cure -- and quickly. Normally drug... approach is to discover key human proteins that can be targeted (hopefully by existing therapeutics that...these experiments, there's so much to do that we can use the whole science community's help. You could... but one are in a pLVX vector background, so you can package them into lentiviral vectors if you need ...FedEx guy has been the best: Thanks, Todd! As you can imagine, it’s also been quite the administrative [email protected] with your order number and so it can be prioritized. ...Arabia -- as well as closer to home in the US and Canada. In fact, we received so many requests (175 and...
  4. Hot Plasmids - December 2019 - CRISPR pooled library, LipoGlo, CRISPRa, and Voltage Imaging Tools

    Type
    Blog Post
    ...genome. Thus the gRNAs responsible for a phenotype can be identified by next-generation sequencing of the...readout. This retroviral mouse CRISPR pooled library can be used for screening of cells that are refractory...lipoproteins through the circulatory systems, but can also lead to the formation of atherosclerotic plaques...transmit electrical information. Such voltage imaging can be performed in intact animals and offers many advantages...found several voltage biosensors and conditions that can be used to monitor real time voltage changes in worms...-mOrange and MacQ-mCitrine. All of these sensors can detect action potentials in worms with fluorescence...libraries was not possible. Primary murine T cells cannot be transduced by lentivirus and have been difficult...
  5. We're Updating Our Hold for Publication Policy

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    Blog Post
    ...publishing research papers can be long and unpredictable. Fortunately, you can submit plasmids at any point... your plasmids! You can make your deposit available before publication and we can update the plasmids’...is posted in order to support these goals. And we can update your plasmids’ citation to the final journal...unresolved sequencing issues or if you’ve asked to cancel the deposit due to experimental problems, legal...article is published. As much as we’d like to, we can’t keep up with every paper that comes out! So, before...
  6. Interview with Bart Newland, Addgene’s Newest Board Member

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    Blog Post
    ...attorneys with a biology background were in demand. Q: Can you tell me a bit about your job currently? What ... the business world from the mindset of a lawyer can bring a new perspective to an organization. I'm used...an organization. I think these are things that I can bring to Addgene -- that comfort with saying, “if... and judgement to the organization and see how I can help. Q: What do you enjoy doing outside of work?...
  7. A Needle in a Base-Stack: Cas9 Structural Biology

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    Blog Post
    ...the CTD can get cozy with DNA. S. pyogenes Cas9 recognizes the PAM sequence 5’-NGG-3’, where N can be any...for genome engineering, and studying its structure can help us understand the parameters of CRISPR experimental...Other techniques, like electron microscopy and FRET, can offer more information on these regions.   Cas9’s...reflecting the less stringent recognition mechanism that can tolerate mismatches in this region. The crystal structure...any base. In Figure 5, we can see why: two arginines make base-specific hydrogen bonds to the guanines...paired to each other? At this point, the protein can switch between the closed and open conformations ...with the rest of Cas9 and is highly flexible; it can rotate about 140 degrees between inactive and active...
  8. Plasmids 101: Dimers and Multimers

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    Blog Post
    ...depicted on the left) is ~5.8 kb in length. The plasmid can exist on as a dimer, depicted on the right, with ...bacterial cells compared to monomers, although this can be prevented via antibiotic selection (Crozat, 2014...instances, please note that the supercoiled dimer band can sometimes overlap with the band corresponding to ...Electrophoresis (CGE) analysis Another technique that can detect multimers is capillary gel electrophoresis...laser-induced fluorescence (CGE-LIF) for detection and can provide data on the quantity of various topologies...strain that is deficient in recombinase protein RecA can reduce the frequency of multimerization, and in some...temperature such as 30°C or optimizing the incubation time can help maintain cultures in log phase growth. In a ...
  9. Transferable Skills Guide: Teamwork

    Type
    Blog Post
    ...expertise and skills of many of my colleagues, I can also orchestrate the formation of high-functioning-but-temporary...to have their own specialties. This type of team can exist indefinitely. If you’re looking for this type...biostatistician, and that one golden-handed technician who can do a certain difficult assay with her eyes closed...the skills are not cherry-picked. We take what we can get since participation on such a team is strictly...without such an association, start one! These teams can offer a sense of solidarity you’re going to crave...about this.” Sometimes the common roles of a leader can be successfully doled out to multiple people: someone...people actually like meetings, but run well, meetings can be a critical part of any successful team. For reasons...
  10. Replacing Paper: Tips for Choosing an Electronic lab Notebook

    Type
    Blog Post
    ... functionalities, it can be difficult to figure out all the ways a given ELN can make your lab life easier... full activity on a project. Can you meet the ELN support team? We cannot stress enough how important ... It is important to know in which ways your data can be exported or printed from the ELN when needed. ... reporting on a project – printing your data out can be required every once in a while, so it is good ...your team. Do you need to make sure your research can be patented? I.e. is automated time stamping (detailed...the times and dates of your actions. These records can take the form of written comments, added files, notes...easier. If you can talk to an ELN support team directly, they should be able to walk you through your ...
  11. Plasmids 101: Stringent Regulation of Replication

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    Blog Post
    ...to be low copy number. Conversely, plasmids that can initiate DNA replication independently are said to...AT-rich regions of DNA. In a test tube, the DNA helix can be separated by DnaA alone, as long as there are ...occupied by ATP-bound DnaA, the proteins cooperatively can separate the DNA strands. Created with BioRender.... available to bind DnaA. Fun fact! As far as we can tell, DnaA doesn’t need to hydrolyze ATP to do its...from the population, while replicating too quickly can slow cell growth or even kill the host cell. For ...replication of pSC101 depends on host-encoded proteins, it can be regulated indirectly via the same mechanisms that... to keep replication in check, high copy numbers can allow RepA to link iterons from two different plasmids...
  12. Four Base Editing Reporters to Monitor and Enrich Editing in Real-time

    Type
    Blog Post
    ...results in two DSBs that can be repaired by NHEJ restoring mCherry expression. Cells can then be sorted using...editing event. GFP positive cells can be sorted and have a significantly higher frequency of base edits ... created an array of base editing reporters that can help you do just that. Monitoring APOBEC and Cas9...due to a frameshift. Restoration of fluorescence can only occur through editing by a APOBEC-Cas9n-UGI ...Fluorescence-activated cell sorting (FACS) and the editing efficiency can be easily determined by comparing cells expressing...editing event would result in GFP fluorescence that can be visualized and sorted via flow cytometry. ...mutated start codon to initiate protein expression, it can be used for an array of detectable proteins including...
  13. Plasmids 101: E. coli Strains for Protein Expression

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    Blog Post
    ... recombinant protein expression. Many challenges can arise when over-expressing a foreign protein in E...restriction/methylation system. This means the strain can neither restrict nor methylate DNA. dcm: Similarly...works well for most experiments, the lac promoter can “leak,” meaning that a low level of expression exists...mostly a problem for toxic protein products, which can prevent the culture from reaching the desired density...any) protein expression? Low expression outcomes can result from variety of sources, so fear not—there...not—there are a few simple troubleshooting measures that can help get you back on track: Compatibility: Double-check...Growth Media: Changing media is tricky, because there can be a trade-off between growth rate and protein quality...
  14. A Modular, Synthetic Origin of Replication

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    Blog Post
    .... Theoretically, large number of compatible ORIs can be created this way with orthogonal regulatory RNAs...constraint. Because the SynORI plasmids are modular, we can swap elements to dial copy number up or down across..., namely transcription factors and riboswitches, can be used to dynamically control copy number. Eliminating...family also suffer from a practical headache: they can form multimers, which reduces stability and makes...incompatibility groups, and members of the same group cannot be used in the same cell at the same time. Starting...
  15. Addgene Stands with the Scientific Community

    Type
    Blog Post
    ...information. Our agility as a nonprofit resource can prove to be vital during this period of constrained...isn't lost. Universities and research institutions can turn to Addgene as a trusted partner for archiving... difficult to maintain or access. These services can be listed as direct costs from a CRO in grant applications...commitment to collaboration and resource sharing, we can navigate this difficult period together. The future...
  16. Download Science-Themed Zoom Backgrounds from Addgene

    Type
    Blog Post
    ...COVID-19 and coronavirus collection page, where you can find plasmid tools to accelerate your COVID-19 research.... Download the blue plasmid background.     You can find all of the plasmids that Addgene offers by visiting... visiting our collections page. Here, you can browse plasmids by field, organism, research technique, ...from Addgene are shipped as bacterial stabs? You can learn about what format your material will come as...
  17. Split Fluorescent Proteins for Studying Protein-Protein Interactions

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    Blog Post
    ...Bimolecular Fluorescence Complementation (BiFC). It can provide a qualitative or quantitative readout of ...of the interaction in vivo or in vitro, and can be used for measuring protein expression or localization... When Protein A and B interact, the FP fragments can assemble the full structure of the fluorescent protein...from a different base FP (for example, EBFP2(1–10) can be used with GFP(11)). Some of the articles linked...color labeling with exogenous fluorophores, which can even be bright enough for single-molecule tracking...interest. Remember that the FP fragments themselves can sometimes interact in cells and produce background...termini are important for their interaction. You can read our previous blog post for more on BiFC principles...
  18. Rabies and Neuronal Tracing

    Type
    Blog Post
    ...this post, I’ll describe how rabies virus (RABV) can be used in the brain to visualize how neurons are... this, cell type specificity of the starter cell can be achieved by using EnvA-pseudotyped RABV, and only...in user-defined cells. For example, a researcher can deliver Cre-dependent TVA to Cre-expressing neurons...it is supplied in trans. Monosynaptic RABV spread can be achieved by modifying the wild-type RABV genome...Wickersham et al., 2007). Thus, RABV transsynaptic spread can be controlled by selectively supplying the G protein... Once in first order neurons, the G-deleted RABV can divide and replicate (since these processes are mediated...RABV monosynaptic tracing is a powerful tool that can be used to reveal brain circuitry, and continue to...
  19. A New Generation of Adenine Base Editors Improves Editing in Primary Human Cells

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    Blog Post
    ...mediate A•T-to-G•C base changes (Figure 1), but it can be challenging to make these base changes, especially... in action they bind DNA so that the TadA domain can edit the short single-stranded DNA stretch that is...catalytically impaired D10A nickase from ABE7.10 can be a problem. Therefore, the authors created ABE8...protospacer adjacent motif (PAM) made up of 5′-NGG-3′ (“N” can be any nucleotide base, followed by 2 guanines). ... in primary human cells  Cell therapy approaches can use base editors to correct mutations in stem cells...which cause sickle cell disease and 𝛽-thalassemia, can be mitigated with the expression of fetal hemoglobin...plays a role for off-target editing effects. ABEs can be introduced into cells as a plasmid or as mRNA ...
  20. Antibodies 101: Isotypes

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    Blog Post
    ...places they can’t reach on their own. The antibody’s heavy-chain constant domains, or Fc region, can be bound...isotypes. Binding to complement system proteins, which can initiate a cascade that helps attack extracellular...useful to know the isotype of an antibody so you can select an appropriate isotype control. An isotype...There are two IgA subclasses: IgA1 and IgA2.  You can learn more about isotype classes in our Antibodies... constant domain gene cluster. DNA recombination can delete intervening heavy-chain constant domain genes... antibodies Read about other techniques that you can use antibodies for in our Intro to Antibodies post...
Showing: 541 - 560 of 1082 results