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We narrowed to 1,018 results for: ple

Showing: 541 - 560 of 1018 results

  1. Viral Vectors 101: Types of viruses

    Type
    Blog Post
    ...that use reverse transcriptase to synthesize complementary DNA copies of their genome upon infecting a ... These viruses have a cargo capacity of ~ 8 kb, ample room for most experimental needs, and can transduce...administration, such as in mouse experiments. AAV also has multiple serotypes (12 naturally occurring and more synthetic...enough for many applications, but in some cases multiple AAVs (co-infection), an alternative virus, or ...applications these viruses are good for, we have plenty of information on that too. May your titer always...

  2. Viral Vectors 101: Viruses as Biological Tools

    Type
    Blog Post
    ...applications, while electroporation of a human, for example, isn’t a reasonable option. Since viruses used...used in labs are derived from viruses that infect people in nature, extra safety measures, like replication...therapies have already been approved in the US for multiple diseases, including hemophilia B and spinal muscular... immunodeficiency virus type 1 preintegration complexes.” Proc. Natl. Acad. Sci. U.S.A. 89 (1992) 6580... Multiple Authors...

  3. AAV Q&A with Tim Miles

    Type
    Blog Post
    ...somewhat more reliable quantitation. Differences in sample treatment across methods can affect reported titer...that users keep titering method consistent across samples within a project. Targeting Your AAV Q4: Are there...for very high MOI on top of very high coverage. Please see this paper for more information about PHP.eB... will very likely cross-react with each other. Multiple labs have tried to transfer PHP.eB's enhanced ...which may be linked to epigenetic changes, such as multiple sclerosis?  A: Basic science and preclinical researchers...

  4. Sweating the Small Stuff: Details in the Lab

    Type
    Blog Post
    ... For example, setting up your pipette tips to match your plate layout can help you track samples as you...change variables to fix a failed experiment.  For example, I used to constantly underestimate time needed... and make a checkmark next to each step as you complete it. Yes, this can be incredibly tedious, but it...remember what's been added and what hasn't. Developing simple habits like these can consistently reduce errors...

  5. Plasmids 101: Methylation and Restriction Enzymes

    Type
    Blog Post
    ... Methylases involved with these processes (for example Dam and Dcm methylases) are independent from the...conversely, activating enzymes such as DpnI. For example, cleavage by XbaI may be blocked due to methylation...those sites will be methylated. Using the same example plasmid as above (which has the XbaI site blocked...methylation by altering your choice of bacteria. For example, if you must use a restriction site that will be...produce DNA that is unmethylated at those sites. Please keep in mind that dam-/dcm- strains may have an...

  6. How to Keep a Lab Notebook for Bioinformatic Analyses

    Type
    Blog Post
    ...labels, since I often work on multiple projects in a given day. For example, instead of naming a file “output...experiments can be found in each folder. This can be a simple .txt document. Record the name, date, and location...folder: You will often use the same data files for multiple analyses. Rather than copying these files to your...result was, so try not to do this. These are the simple rules that I try to follow in my own research. ...

  7. Luminescent Imaging with Nano-lanterns

    Type
    Blog Post
    .... This luminescent technology is based on the principle of bioluminescence resonance energy transfer (...photobleaching/phototoxicity, they successfully imaged multiple subcellular compartments over several minutes ...improve long-term imaging, which is limited by depletion of luminescent substrate, they synthesized diacetyl... handy. This approach is especially suited to complement optogenetic tools, where light used for excitation...external light source. Therefore, especially thick samples may not be appropriate for use with Nano-lantern...

  8. Why and how to start studying chromatin (with Addgene's help)

    Type
    Blog Post
    ...interact with each other. Chromatin regulation is complex and influences--directly or indirectly--many other... we definitely don’t have that you really want, please let us know! And if you study chromatin and want...t see the gene you’re looking for on this list, please consider searching directly on our website. ...phosphoinositol-dependent binding of the SWI/SNF-like BAF complex to chromatin after T lymphocyte receptor signaling...not the first Addgene blog post about chromatin--multiple Addgene colleagues and guest bloggers have written...

  9. Early Career Researcher Toolbox: Social Media for Scientists

    Type
    Blog Post
    ...help my research and my career. Here are three examples of how using social media has helped me or my ... can start using Twitter and provide real-life examples of how they and others have used Twitter professionally...also find Dr. Page and Hello PhD on Twitter: @ThePurplePage and @HelloPhD "Role" call. #BLACKandSTEM what...do you do? — Stephani Page, PhD 👩🏾‍🔬💜 (@ThePurplePage) February 13, 2014 2. "The Scientist's Top... for Twitter pros as well, like tip #4: “Follow people for about a week and then unfollow those who do...

  10. The time and cost required to make a plasmid

    Type
    Blog Post
    ...outlined in Table 1.   Of course, this is for the simple insertion of a PCR product into a vector. The time...calculate, in both our 2018 and 2023 poll, most people said it took them more than 22 hours, so at least...published in. That’s also a great way to get to know people doing interesting and relevant work in your field...That way, you can run one PCR, one gel, etc. for multiple plasmids at once. Over time, this can save you... Multiple Authors...

  11. CRISPR 101: Cas9 vs. The Other Cas(s)

    Type
    Blog Post
    ...is guided by an RNA molecule, the gRNA, to a complementary site in the genome where it will proceed to ...degradation. Cas3 DNA digestion can proceed up to multiple kilobases, but only in one direction. Cas3 is ...that Cas14 also indiscriminately cleaves non-complementary ssDNA when engaged by its target sequence. Why...used in lieu of siRNA or shRNA for efficient, multiplexable RNA knockdown. Cas13 has also stirred up interest... the other Cas family members make for useful implements in the scientist toolbox. If your genome engineering...

  12. Transferable Skills Guide: Problem Solving

    Type
    Blog Post
    ...ill-defined. In this article, I try to clarify what people mean when they ask about “problem solving skills...problem.  Step Three: Decide on a strategy and implement it.  Step Four: Assess how effective your strategy...the cause. To test the most likely cause, you implement an experiment designed to fix the problem. Then...modifications, and ultimately decided on the changes to implement. As with any skill, there will always be room ...Know your strengths! Research experience presents ample opportunities to practice problem solving skills...

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

    Type
    Blog Post
    ...conduct research that fails less and helps more people, we need to take sex into account. Gendered Innovations...s Health Initiative Hormone Therapy Trial, for example, found that the study saved lives. Reduced use ...analysis and 2) provides case studies with concrete examples of how gender analysis leads to discovery and ...experiments—which can lead to failed research. For example, an international research team from Norway and...studies that draw from a body of research that is incomplete (because sex was not analyzed) may harm humans...

  14. RANbodies: Reporter Nanobody Fusions

    Type
    Blog Post
    ...of the sample due to the smaller size of RANbodies compared to antibodies. Staining with multiple HRP-RANbodies...used at a time. Samples can be sequentially stained with two HRP RANbodies if the sample is stripped. HRP-RANbodies...process that’s difficult for individual labs to complete. The nanobody based RANbody platform from the ...

  15. Troubleshooting Your Plasmid Cloning Experiment

    Type
    Blog Post
    ...Assembly, Golden Gate etc.), and try to find the simplest way to do it (i.e. avoid assembling too many fragments...methylation like XbaI, ClaI, etc. If you are doing multiple digestion be sure that the buffers and temperature...Remember to quantify the DNA concentration of all the samples by gel or by nanodrop to assemble the  fragments...molar ratio. Assembly of the fragments There are multiple ways you can assemble the different parts of a...plasmid. Many vectors contain sequences that are complementary to standard sequencing primers. If not, design...

  16. Plasmids 101: CcdB - The Toxic Key to Efficient Cloning

    Type
    Blog Post
    ...ccdB aspect, Gateway takes advantage of the same principle that cells will not propagate while expressing...system contains ccdB. A successful insertion will completely replace ccdB with the investigator’s insert of...interest.   CcdB resistant E. coli strains complete the system But ccdB alone isn’t the only key to...most appropriate method for your own experiments. Please check out Addgene’s Choosing a Molecular Cloning...protein involves poisoning of DNA-topoisomerase II complexes.” J Mol Biol. 1992 Aug 5;226(3):735-45. PubMed...

  17. Design Tips for Prime Editing

    Type
    Blog Post
    ...extended to include a primer binding site (PBS) complementary to the genome and a reverse transcriptase (RT... the experiment. Now, onto the tips! General principles pegRNA design If you’re just starting out with... nicking the newly synthesized strand before heteroduplex resolution, which can lead to indels (Anzalone...edited strand as a template during DNA repair. Test multiple nick sites, starting with sites about 50 bp upstream...Nureki, O. (2014). Crystal Structure of Cas9 in Complex with Guide RNA and Target DNA. Cell, 156(5), 935...

  18. 6 Tips for Grant Writing

    Type
    Blog Post
    ...bit of advice I can suggest, is to give yourself plenty of time when writing a fellowship proposal. Yes... you learn new techniques and interact with new people. If you are looking to move to a different university...there is an increasing realization that, if we want people to keep contributing to science, it would be helpful...audience. My lay paragraph typically includes a couple of sentences of introduction to my area, a line...great information on ELISA troubleshooting and multiplex assays.   Additional Resources on the Addgene ...

  19. Savvy Advocates Needed to Navigate a Scientific Enterprise in Flux

    Type
    Blog Post
    ...challenges and opportunities for us all.  For example, implementation and compliance for the FLSA changes, including...turn. The first is the passing and upcoming implementation of the Federal Labor Standards Act (FLSA) which...remain mixed in their opinions on the subject – for example, Yale graduate students filed for union elections...challenging, but will be aided by the development and implementation of new strategies designed to cope with current...

  20. Adeno Associated Virus (AAV) for Cell and Gene Therapy

    Type
    Blog Post
    ... the mutation or replacing the gene. A notable example is the treatment of Severe Combined Immune Deficiency...normally functioning T cell repertoire (1). In this example, it was possible to transduce the T-cells outside...based delivery techniques may hold the key. In the simplest form, translating gene therapies from conceptual...already have ways to prevent AAV infection. For example, prevalence of neutralizing antibodies against ...: 14566358.  2. Hirsch, M. L., et al. "Self-complementary AAV mediates gene targeting and enhances endonuclease...
Showing: 541 - 560 of 1018 results