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We narrowed to 957 results for: omp

Showing: 161 - 180 of 957 results

  1. Antibody Validation Using the Indirect ELISA Method

    Type
    Protocol
    ...science. General Considerations The ELISA can be completed in a single day or broken up over several days...ELISA plate washer. Equipment Spectrophotometer compatible with 96-well plates 1–10 µL single channel pipette...

  2. Kit Free RNA Extraction

    Type
    Protocol
    ...to allow for dissociation of the nucleoprotein complexes. The effectiveness of your RNA isolation will ...

  3. Making CRISPR Plasmids Using Fragmid

    Type
    Blog Post
    ...plasmid. It shows you each component, with the ID of the component plasmid, component name, size, and any comments...module, are the components. Using the handy website, users can simply select the components they want for...able to select components that work with your design. Note that you can browse all components by clicking...Fragmid doesn’t have the components I want? You can make your own Fragmid components! Ensure your backbone... Each module of Fragmid represents a class of components, such as your CRISPR enzyme or Pol 2 promoter...’ terminus, and selection markers. Need fewer components? A spacer can be inserted instead, allowing everything...everything to stay in the correct order. Most Fragmid components are available for request on the Addgene website...

  4. 27 Hot Plasmids from 2016

    Type
    Blog Post
    ...easily design, generate, and test constructs composed of complex genetic elements. To demonstrate the usefulness...several GMAP-compatible vectors for a wide variety of biological applications. First, GMAP-compatible backbones...mammalian systems. The DULIP system is comprised of Gateway-compatible plasmids in which “bait” proteins are...plasmids designed to complement unused auxotrophies in Saccharomyces strains by compensating for histidine (... do so have been hindered by a lack of tools, compounds, and assays to monitor activation of these receptors...including a stop codon) formats. The collection is compatible with both standard Gateway cloning and the FNLCR...assembly platform for generating both simple and complex DNA constructs Have you ever wished you could ...

  5. Truncated gRNAs for Regulating Gene Expression

    Type
    Blog Post
    ...DNA strands complementary and non-complementary to the gRNA, respectively. When complexed with a truncated... (nuclease-competent) Cas9 interacting with either a full-length gRNA (20nt complementarity to target ...) or truncated gRNA (≤15nt complementarity to target site). When complexed with a full-length gRNA, native...with which it is complexed rather than the protein itself. When native Cas9 is complexed with a gRNA that...transcriptional activator to the Cas9-gRNA complex. When this complex is directed to a target upstream of the...Figure 3: Gene repression with nuclease-competent Cas9. When complexed with a truncated gRNA, Cas9 is still... directed to virtually any genetic locus when complexed with an appropriately designed small RNA, or guide...

  6. Viral Vectors 101: Viral Vector Elements

    Type
    Blog Post
    ...replication components are called helper plasmids. Below, we’ll breakdown the vector components first by ...nucleocapsid viral components needed for viral packaging, assembly, and display. Pol, the last component, contains...there are bonus components to these vectors that aren’t reflected in the common components outlined above...lentiviral guide outlines these components.     Fig. 1 - Viral vector components of retrovirus  Adeno-associated...production reasons, multiple plasmids with many components are used to package lab grown viruses. It can... protocols! In reality, there are many shared components across viruses (AAV, lentivirus, etc.) and their... what gag and pol stand for, we demystify all components of viral vectors here! Need Virus? Check out...

  7. Finding Your Perfect Job After University

    Type
    Blog Post
    ...role at a commercial virology testing company. This particular company carries out clinical trials of drugs...treatments for external companies. It was really exciting to work in a larger commercial company; the pay was good...the company and that my ideas and changes were welcomed, which is a benefit over larger companies which...obtaining work in scientific research would be very competitive, and so I applied for a summer role as a Research...This was particularly interesting as I had just completed my thesis on the study of e2Fb, which is a tumour...without issues. Because the primary purpose of the company was to fulfill contracts, my work sometimes felt...strong point. Working in industry: virology On completing my MSc, I again had to look at what type of work...

  8. Viral Vectors 101: Voltage Indicators

    Type
    Blog Post
    ...different components, but all contain a FRET pair, a genetic component, and a small molecule component. Early...developed voltage sensor with a genetic component. The VSD components have historically come from a variety...have been engineered to have kinetics comparable to its one component counterpart and is significantly brighter...cellular localization component is typically handled by either the fluorescent component (i.e., when the dye...tools have two main elements: a voltage-sensing component and a fluorescent output. The GEVI family of gadgets... a hybrid approach and utilize both a genetic component as well as a small molecule which we will describe...reporter and the mechanism of their voltage sensing component, which separates them into two distinct categories...

  9. Typing CRISPR Systems

    Type
    Blog Post
    ... as Cascade (CRISPR-associated complex for antiviral defense) complexes, while Class 2 use single protein...effector complex after binding to a target. Class 1 is the most abundant of the two, comprising about 90%...tools using a Cascade complex is tricky, as you need to ensure every unit of the complex expresses at similar...have similar compositions and mechanisms, with the biggest differences being the components present in ...in the Cascade complex. Figure 2: Schematic of a Cascade complex (blue subunits) with a guide RNA...primary effector complex but are defined by the accessory Cas proteins in the Cascade complex. Despite a tenuous...the Cas effector may have the same name but be completely different! They are merely linked by a similar...

  10. CRISPR 101: Mammalian Expression Systems and Delivery Methods

    Type
    Blog Post
    ...are compatible with all your components. Next, consider the best form for those CRISPR components based...introducing those CRISPR components into your target cells. Delivery of these components into mammalian cell...most effectively achieved with different CRISPR components. Select the appropriate CRISPR tools for your...will you be editing? Do you want to deliver the components as DNA, or would mRNA or protein delivery be ...of the first steps is to identify what CRISPR components you will need to deliver. Minimally, one or more... the best method for introducing those CRISPR components into your target cells.  Mammalian CRISPR expression...best practices for efficient delivery of CRISPR components. If you are new to CRISPR or your model system...

  11. Antibodies 101: Flow Cytometry

    Type
    Blog Post
    ...information about the cell’s complexity. A cell with a high degree of internal complexity such as extensive membranes... prepare sets of controls to aid with compensation. Compensation is the process of correcting spectral...need to analyze a specific subset of cells in a complex population. Have you spent hours in the biosafety...cell’s size, granularity, and target protein composition. It is used for a variety of applications such...and detecting specific populations of cells in a complex tissue or sample (1). When combined with cell sorters...instrument collects information about the cell’s size, complexity and fluorescence intensity.   What flow cytometers... cytometers detect Cell size and complexity, measured using visible light scatter The optical system has...

  12. Working with Nuclear Receptors

    Type
    Blog Post
    ...co-coregulator) partners. The coregulator ‘core’ complex is composed of a tightly-bound invariant group of proteins...coregulators exist as multiprotein complexes, a member of a single coactivator complex can serve as a rate-limiting...seeking to compliment these consortium-generated datasets with a systematic effort to compile NR- and coregulator-related...physiological processes are controlled by deliberate and compulsory actions instigated by the central nervous system...factors found in metazoans. The superfamily is comprised of 48 human and 49 murine members. NRs permit ...metabolic, and immune response programs. NRs also encompass one of the most successful targets for drugs currently...interact directly with NRs) exist in steady-state complexes with secondary coregulator (or co-coregulator)...

  13. Degrading DNA with Cascade-Cas3

    Type
    Blog Post
    ...different compositions of the Cascade complex. Created with BioRender.com.   The Cascade complex Cascade... an important component of the CRISPR Cas3 system. Most Cascade complexes are comprised of a combination...systems work using a multi-Cas complex called Cascade (CRISPR-associated complex for antiviral defense). Cascade... Cascade complex from CRISPR type I-E. Created with BioRender.com. Thanks to this complex, developing... It can be difficult to ensure comparable expression of all components of Cascade for maximum efficiency...Cascade-Cas3 plasmids in Addgene's collection! A "complex" system Cas3 is part of the Class 1 CRISPR family...from different species and will have different compositions of Cascade. Although seemingly ubiquitous in...

  14. Delivery Methods for Generating iPSCs

    Type
    Blog Post
    ...iPSC cell lines, which can complicate comparisons made between lines. Incomplete silencing of transgenes ...are prone to incomplete silencing of reprogramming transgenes, which leads to incomplete reprogramming... heterogeneity between clones. This can complicate comparisons made between samples, i.e. transgene expression...efficiency of reprogramming when compared to other methods, even when compared to integrating viral approaches...less time to complete. Morphology is a simple and fast to evaluate. iPSCs should form compact colonies that...and have a higher efficiency of generating iPSCs compared to other delivery methods. Retrovirus has a cloning...lentivirus has similar rates of reprogramming when compared to MMLV-derived retroviruses, it is not as well...

  15. Experimenting in an Open Source lab: from CRISPR to Cats

    Type
    Blog Post
    .... The team at the Victoria Makerspace is composed of computer scientists, engineers, and biologists working... elegans as a computer model. When I first heard about it, I got excited from a computer scientist perspective...defects above 25 degrees. Computer scientists and biologists work together to compare the predictions about...interests in biology and computing. In 2009, he founded a team at UVic to compete in iGem, an MIT-sponsored... really epitomizes what the makerspace is. The computer scientists can get involved in modelling, the ...Developing these skills relies upon biologists and computer scientists learning and communicating with one...coding the molecular biology of the worm into a computer model that recapitulates how molecular biology...

  16. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    ...of oxidation (dehydrogenation). MitoTimers are composed of a Timer protein fused to the mitochondrial ...upregulate an alternative pathway or induce other compensatory mechanisms. How can protein function be dissected...’s lab, acutely depleted proteins from their compartment of action via sequestration. The strategy relies...outer mitochondrial membrane. Sequestration is accomplished by knocking down your endogenous protein of ...May 1;126(Pt 9):2102-13. Assemble multiple component plasmids from building blocks via Golden GATEway...GATEway Cloning Frustrated by the complexity of assembling recombination of transgenesis constructs? A new...cloning kit simplifies the cloning process for complex DNA constructs, particular for those involving ...

  17. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ... pre-mRNA splicing in a compound screen. Their reporter system is comprised of two plasmids: Luc (intronless... Vacuolar Compartment mCherry-Rab5 BFP-Rab5 GFP-Rab5B Vacuolar & Budding Compartment GFP-Rab4B ...insights into the make-up of multiprotein complexes like the EPC1 complex.. Going one step further, they also...about it! Split Cas9 system The Cas9 protein is composed of an n-terminal DNA recognition domain and a ...sequencing or other readout. Cellular memory is accomplished by the use of sequence-specific enzymes (e.g...calculated (the BRET ratio), which allows for compensation of varying fusion protein expression levels....more powerful LID, the authors recently used computational protein design, phage display and high throughput...

  18. Plasmids 101: Restriction Cloning

    Type
    Blog Post
    ...important that the digest go until completion. The time required for complete digestion varies for different...want to use electro-competent cells instead of the more common chemically-competent cells. You should perform...fragment of DNA (insert) that has been cut by compatible restriction enzymes. An enzyme, DNA ligase, then...plasmid to the new fragment thereby generating a complete, circular plasmid that can be easily maintained... are cut with restriction enzymes to generate compatible overhangs that allow them to bind. Ligase is ...your backbone and insert - these both must have compatible cut sites for restriction enzymes that allow ...    Alternatively, this whole process can be completed using a single enzyme if your insert is flanked...

  19. Career Insights: Technical Support Specialist

    Type
    Blog Post
    ... work for biotechnology companies and help other researchers use the company’s products successfully. ...guidance, and resolve complaints about products. As you might expect, different companies produce different... duties. Bigger companies have dedicated technical support teams, sometimes comprised of several layers...cells (manufactured by one of the larger biotech companies), and as far as I remember, it was the first time...time that I was absolutely convinced it was the company’s fault. I contacted them to express my concerns...rather painful to get there: I had to send the company detailed documentation of my results, answer loads...takes some time (several years) to build up comprehensive lab experience. With this valuable training ...

  20. Plasmids 101: E. coli Strains for Protein Expression

    Type
    Blog Post
    ... to accommodate high protein levels including:  ompT: Strains harboring this mutation are deficient in...proteins. lon protease: Strains where this is completely deleted (designated lon or Δlon) similary reduce...IPTG-inducible strain containing T7 RNAP (DE3) F- ompT lon hsdSB (rB- mB-) gal dcm (DE3) General protein...vector cannot have p15A origin of replication F- ompT lon hsdSB (rB- mB-) gal dcm (DE3) pLysS (CamR) ...vector cannot have p15A origin of replication F- ompT lon hsdSB (rB- mB-) gal dcm (DE3) pLysE (CamR) ...which results in longer transcript half-life F- ompT lon hsdSB (rB- mB-) gal dcm rne131 (DE3) General...RNAP; IPTG may still be required for expression F- ompT lon hsdSB (rB- mB-) gal dcm araB::T7RNAP-tetA General...
Showing: 161 - 180 of 957 results