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We narrowed to 850 results for: NTS

Showing: 421 - 440 of 850 results

  1. CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More

    Type
    Blog Post
    Published
    Sept. 28, 2020, noon
    ...promote homologous recombination of linear DNA fragments. Since recombineering does not contain a selection... in the Cas9 or spacer sequence, and these rare events are easily detectable using PCR. The system also...can be also linearized with XbaI to insert extra elements, like a repair template, using Gibson Assembly...system called INTEGRATE (Insertion of Transposable Elements by Guide RNA-Assisted Targeting) and the Zhang...transposition. INTEGRATE consists of four major components including (1) a CRISPR RNA, (2) four proteins...transcription factors, phage proteins, and RNAP components (Dong et al., 2018). They found a transcription...

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

    Type
    Blog Post
    Published
    July 8, 2025, 1:15 p.m.
    ...structure you want to achieve before starting your experiments. Don’t dive directly into the pipetting! MoClo...be used directly in your assemblies. Preparing reagents Keep it cool You’ve done all the learning and ...What really matters is the molar ratio of your fragments. To increase your ligation efficiency, we recommend...Calculation for the plasmid volume for MoClo experiments. The term ‘insert’ refers to plasmids carrying... but they also work with very small volumes of reagents which lowers experimental costs. If you are planning...

  3. Plasmids 101: Shuttle Vectors

    Type
    Blog Post
    Published
    May 20, 2025, 1:15 p.m.
    ...’s convenient for researchers who want to do experiments in complex eukaryotic cells, but would prefer...resistance gene, so it isn’t a good fit for all experiments. And the list of antibiotic options doesn’t end...Auxotrophic selection is especially common in yeast experiments because yeast tend to spontaneously develop resistance... but it isn’t common for simple transfection experiments. Reporters Finally, in some cases a researcher...gene they simply can’t turn on! Wait, what even counts as a shuttle vector? Are viral vectors shuttle ...

  4. Transferable Skills Guide: External Collaborations

    Type
    Blog Post
    Published
    April 19, 2018, 1:25 p.m.
    ...methods and equipment between lab groups and departments. You don’t have to get these types of projects...discipline. Getting companies to come to these types of events can be very tough. First I needed to find the right...-to-date on your collaborators’ professional developments without directly contacting them. One great ...professional updates. Whenever I see interesting events, articles and newsletters I make sure to share ...Reading "Collaborate: An Imperative for Graduate Students" from Inside Higher Ed Additional Resources on...

  5. CRISPR Activation: A Practical Guide

    Type
    Blog Post
    Published
    Aug. 18, 2016, 2:30 p.m.
    ...upregulation of essentially any gene by fusing mutants of Cas9 deficient in DNA cutting activity to a ...cannot help you pass. In addition, activation experiments often require quite a bit of tuning before you...wide variety of activators you can use for your experiments. We have found that SAM (Konermann et al., 2014...be true given the results of previous RNA-seq experiments (Konermann et al., 2014, Chavez et al., 2014,...works reasonably well.  Best of luck on your experiments!  Many thanks to our guest bloggers Marcelle...

  6. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

    Type
    Blog Post
    Published
    Oct. 19, 2017, 1:54 p.m.
    ...Cre-lox or one of the FLP-FRT systems in your experiments. Mutated FRT and loxP sites in Flex Vectors –... – inversion only once All recombination events mediated by FLP or Cre are reversible. Whereas the excision...individual neurons one could reduce the plasmids amounts to reduce transfection efficiency. However, if ...recombinase expression while proteins needed for later experiments (cellular tracking for instance) can be controlled... inducible Cre versions can be used in these experiments. This elegant combination of Cre, FLP, and inducible...

  7. Plasmids 101: Stringent Regulation of Replication

    Type
    Blog Post
    Published
    June 20, 2024, 1:15 p.m.
    ...produce lower amounts of protein, which can be useful if the product is toxic in high amounts. These plasmids...appropriate time—for example, the cell must have enough nutrients available to complete the entire round of replication... are efficient for the host, as no additional components are necessary. Iterons are not the only tool ...sequence forming this pseudoknot, so its binding prevents the mRNA from folding and blocks RepZ translation...

  8. Hot Plasmids Spring 2024

    Type
    Blog Post
    Published
    May 23, 2024, 1:15 p.m.
    ...recombination and avoid separation of the multiplexed elements. The team designed sgRNAs that include an internal...by this tool as well as additional potential improvements to the editor. Find PE7 plasmids here!   ...in the rhodopsin domain conferred several new improvements. The new variant has increased signal-to-noise...voltage indicator Ace2N-mNeon also led to similar improvements in performance.      Figure 4: Structure...trans-splicing. By splitting a large gene into smaller fragments, delivering them by individual AAVs, and then ...

  9. Plasmids 101: Broad Host Range Plasmids

    Type
    Blog Post
    Published
    April 11, 2024, 1:15 p.m.
    ...origin of replication (ori) contains structural elements that are versatile and adaptable. Variation in...bacterial hosts (Doran et al. 1998). Regulatory elements within the ori also control copy number and stability...differentiated from one another. Based on their structural elements, oris interact with various RNA polymerases and...Host range depends on whether the host has the components to recognize the ori and allow for replication...consist of a plasmid’s ori and all of its control elements, from broad host range plasmids can be used to...

  10. Antibodies 101: Immunoprecipitation

    Type
    Blog Post
    Published
    Dec. 7, 2021, 2:15 p.m.
    ...approaches allow for optimized strategies for differing amounts of target proteins or antibody bonding strengths...target protein or if the target is present in low amounts. In this method, the free antibody is incubated...magnetic beads with commercially available chemical agents. Directly conjugating the primary antibody has ... a non-denaturing buffer containing non-ionic detergents, such as NP-40 or triton-X, to preserve native...than endogenous proteins. Data obtained from experiments using a tagged protein may not be translatable...

  11. Beyond the Blood Brain Barrier: PHP.B Enters the Central Nervous System Via the LY6A Receptor

    Type
    Blog Post
    Published
    July 23, 2019, 12:46 p.m.
    ... by first generating a diverse library of AAV9 mutants that had a small randomized peptide loop inserted...screened in vivo in C57BL/6J mice strains to identify variants that reached the CNS following IV injection. Compared...how the method used to select novel AAV capsid variants could unintentionally limit the utility of candidate...E., et al. "Cre-dependent selection yields AAV variants for widespread gene transfer to the adult brain...research. Visit Our AAV Guide Page for info on AAV components, common uses of AAV, and much more! Find ready-to-use...

  12. New Tools Enable CRISPRa for Neuroscience Applications

    Type
    Blog Post
    Published
    July 2, 2019, 12:51 p.m.
    ...approaches. Here, we’ll highlight these recent advancements and offer commentary on their application to...are needed to deliver all the protein and RNA components, and several sgRNAs must be screened to identify...robust and specific upregulation of the targeted variants without inducing expression of the non-targeted... between the Bdnf I and Bdnf IV overexpression variants were other immediate-early genes associated with...activity, such as Fos, Egr3, and Arc. Thus, CRISPRa represents a powerful method to dissect the individual and...

  13. Designing Your Chalk Talk for the Academic Job Interview

    Type
    Blog Post
    Published
    March 12, 2019, 1:37 p.m.
    ...academic job market? In addition to all of the documents you submit, you will need to present a "chalk .... Given this large sum, one might expect that applicants would work for months to hone a chalk talk. Yet... use the information. In contrast, a list of experiments or a laundry list of molecules to screen has ...is a significant need for healthy livers for transplants. Growth of new livers in culture could resolve...learned and/or now be possible to do? Research accomplishments are remembered for the problem solved, not ...

  14. New Optogenetic Tools for Cytoskeleton and Membrane Control

    Type
    Blog Post
    Published
    Sept. 28, 2023, 1:15 p.m.
    ...Researchers have designed a variety of photoswitchable elements into many other types of proteins. Here, we’ll...sword Microtubules are one of the most important components of the cytoskeleton. They provide cells with ... will surely be useful for a wide variety of experiments. Opto-RhoGEFs manipulate cell morphology and ...phospholipase D (PLD). Now they present a set of variants called SuperPLDs with 10, 30, 50, and 100 times... and cytotoxicity better than the 50x and 100x variants. But those super-high efficiency constructs are...

  15. Treating Muscular Dystrophy with CRISPR Gene Editing

    Type
    Blog Post
    Published
    Jan. 26, 2016, 3:30 p.m.
    ...For such a therapy to be successful, multiple requirements must be met. First, CRISPR would need to be ...desirable. Should stem cell editing occur, the CRISPR components would only need to be expressed for a short period...skipping therapies would be applicable to 80% of DMD patients. Figure 1: Exon skipping strategies for...the technology is advancing, precision editing treatments may also be closer than we might expect.   References...expression in cells from Duchenne muscular dystrophy patients through genomic excision of exon 51 by zinc finger...

  16. The 12 Days of CRISPR

    Type
    Blog Post
    Published
    Jan. 3, 2023, 2:15 p.m.
    ...CRISPR, Addgene gave to me: Functional Genetic Variants Revealed by Massively Parallel Precise Genome ...prime editing systems by manipulating cellular determinants of editing outcomes🎵 On the 5th day of CRISPR...CRISPR, Addgene gave to me: High-fidelity Cas13 variants for targeted RNA degradation with minimal collateral...prime editor with enhanced editing efficiency in plants 🎵 ...

  17. Antibodies 101: Multiplex Immunofluorescence

    Type
    Blog Post
    Published
    May 23, 2023, 1:15 p.m.
    ...consider strategies using other types of affinity reagents like fluorescently-labeled nanobodies or scFvs... label such as horseradish peroxidase (HRP). Experiments using enzymatic labeling can be difficult to ...t forget the controls! The necessary control experiments will vary depending on your specific experimental...design. Like selecting an antibody, some of the requirements are similar to standard IF. Here, we’ll walk...intensity patterns are similar between the two experiments. If not, it might be that the other antibodies...

  18. Plasmid Grant to Aid Cervical Cancer Screening in Ghana

    Type
    Blog Post
    Published
    Nov. 8, 2016, 3:30 p.m.
    ...PCR-based cervical cancer screens. This third grant represents the continuation of our efforts to provide resources...Bengyella and Kwabena Duedu who were awarded similar grants back in June, Gladys will be using plasmids in ...Seeding Labs Plasmid Grant to a third round of applicants at the start of 2017 and would like to thank ...Superhero 5k Find New Cancer Resources Learn About Our Grants to Rare Disease Research Resources on Addgene....

  19. Newly Updated AAV Data Hub!

    Type
    Blog Post
    Published
    Feb. 7, 2023, 2:15 p.m.
    ...community submissions for Addgene and CLOVER AAV experiments - and we've updated our submission process so... are selecting AAVs that will work for their experiments, and a key step for that is sharing the data ... negative data helps others to design better experiments (and wouldn't it be nice if those weeks and months... the Data Hub to help select an AAV for your experiments, let us know ([email protected]) and we might...

  20. FLEx Technology and Optogenetics: Flipping the switch on gene expression with high spatial and temporal resolution

    Type
    Blog Post
    Published
    Oct. 17, 2019, 12:28 p.m.
    ...-stimulate mammalian neurons and alter synaptic events: a new technology was born, and the term “optogenetics...infect defined neuronal subpopulations, and this represents a major downside in optogenetics (Belzung et ... delivery of Cre recombinase in mouse primary afferents." Scientific reports 8.1 (2018): 7321. PubMed ...and Peter Hegemann. "Rhodopsin-regulated calcium currents in Chlamydomonas." Nature 351.6326 (1991): 489...neurons: tools, applications, and experimental constraints." Neuron 95.3 (2017): 504-529. PubMed PMID: 28772120...
Showing: 421 - 440 of 850 results