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We narrowed to 932 results for: omp

Showing: 141 - 160 of 932 results

  1. Ligation Independent Cloning

    Type
    Protocol
    ...restriction digest protocol ). After the digest is complete, you will need to separate the linearized vector... A typical T4 Pol reaction is shown. Mix the components (add the T4 last) and incubate at room temperature...of T4 Pol in the following step. This may be accomplished by gel purification . Step 5: Create Insert ...

  2. Centrifugation

    Type
    Protocol
    ... and using a centrifuge to separate different components in a liquid sample....laboratory equipment used to separate different components in a liquid sample. It achieves this by using... you use, most will have essentially the same components. The photo to the left shows a front view of ...

  3. Gibson Assembly Protocol

    Type
    Protocol
    ...2009). Regardless of fragment length or end compatibility, multiple overlapping DNA fragments can be joined...overhangs by chewing back from the DNA 5’ end. Complementary DNA fragments can subsequently anneal to each... DNA Ligase - covalently joins the annealed complementary DNA fragments, removing any nicks and creating...

  4. Protocol - How to Create a Bacterial Glycerol Stock

    Type
    Protocol
    ...bacterial strain and you will not need to obtain more competent cells and retransform. Bacteria on an LB agar ...streaking onto LB agar will prevent it from thawing completely and will improve the shelf life. It is very important...

  6. Transfection for Recombinant Antibodies

    Type
    Protocol
    ...biosafety level 2. Please ensure that you are in compliance with your institution’s biosafety regulations...tips Pipettor and pipettes Benchtop centrifuge compatible with 50 mL conical tubes Automated cell counter...

  7. AAV Purification by Iodixanol Gradient Ultracentrifugation

    Type
    Protocol
    ...vivo . The iodixanol gradient in this protocol is composed of steps that separate out contaminants from an...exchange and concentration Note: Both steps could be completed in one (long) day. Equipment Ultracentrifuge T70i...

  8. Protocol - How to Run an Agarose Gel

    Type
    Protocol
    ...). Microwave for 1-3 min until the agarose is completely dissolved (but do not overboil the solution, ...room temperature for 20-30 mins, until it has completely solidified. Pro-Tip If you are in a hurry, the...

  9. Handling Plasmids from Addgene - Purifying Plasmid DNA

    Type
    Protocol
    ...bacterial cultures , please see those pages. Several companies, such as (Link opens in a new window) NEB , (Link...but within a smaller volume of buffer that is compatible with the next solution. Add a denaturing solution...

  10. 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...

  11. Kit Free RNA Extraction

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

  12. 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...

  13. 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 ...

  14. 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...

  15. 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...

  16. 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...

  17. 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...

  18. 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...

  19. 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...

  20. 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...
Showing: 141 - 160 of 932 results