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We narrowed to 885 results for: TRI

Showing: 821 - 840 of 885 results

  3. Cloning

    Type
    Guide
    ...used to create recombinant DNA. Restriction Enzyme Cloning Restriction enzyme (endonuclease) based molecular...sites in the DNA. In restriction cloning, scientists utilize specific restriction enzymes to cut dsDNA... find a protocol for restriction cloning and an in-depth breakdown of restriction digests on our website...Empty Backbone Guide Genome Engineering Guide Restriction Enzyme Gateway TOPO Gibson Type IIS Ligation ...reasons, remains one of the most popular today. Restriction enzymes, which are naturally produced by certain...during a ligation reaction with T4 DNA ligase. Restriction enzyme cloning benefits from the hundreds of ...cloning and expression contain several popular restriction enzyme sites. You can easily move (subclone) ...

  4. Molecular Biology Reference

    Type
    Guide
    ...contains several restriction enzyme sites, enabling easy insertion of DNA by restriction enzymes digestion...used for cloning by restriction enzyme digestion. For more information about restriction enzymes check out...human genome. Digesting the human genome with restriction enzymes would yield about two million DNA fragments...plasmid. By the 1970s the combined discoveries of restriction enzymes, DNA ligase, and gel electrophoresis ...antibiotic-resistance gene, and at least one unique restriction enzyme recognition site. These elements allow... not carrying the plasmid. Additionally, the restriction enzyme site(s) allow for the cloning of a fragment...driven by the promoter. As a general rule, the restriction sites in the MCS are unique and not located elsewhere...

  5. Adeno-associated virus (AAV) Guide

    Type
    Guide
    ... therefore non-pathogenic and generally do not trigger a strong immune response. These features have made...with most standard cloning methods, including restriction enzyme, Gibson Assembly, or Gateway. Some transfer...transfer plasmids may have limited restriction sites or may only be compatible with certain cloning methods...are not efficiently transduced by AAV2, and is distributed more widely in the brain. Many of these hybrid...enhanced function after intravenous administration, distributing widely throughout the organism to target large... and Duchenne muscular dystrophy. Many clinical trials of AAV-based gene therapies are underway and are...Cells AAV Purification by Iodixanol Gradient Ultracentrifugation AAV Titration by ddPCR AAV Titration by qPCR...

  6. Antibody Guide

    Type
    Guide
    ...light orange, that bind to a specific antigen, triggering an immune response. The stem of the Y structure...single antibody signal. Here, HRP is used as a colorimetric, in which the reaction produces a colored product...the amount of enzyme present in each well. In colorimetric assays, protein amount can be determined by ..., lyse, and then collect protein extract by centrifugation or affinity columns. Mix samples with a primary...immobilized on agarose or magnetic beads. Using centrifugation (for agarose beads) or a magnet (for magnetic... your samples using permeabilization, antigen retrieval, and/or sectioning. Block to reduce non-specific...tissue sections are assayed and the extracellular matrix remains intact. The signaling molecule may be a...

  7. Adenovirus Guide

    Type
    Guide
    ...with most standard cloning methods, including restriction enzyme, Gibson Assembly, or Gateway. Some transfer...transfer plasmids may have limited restriction sites or may only be compatible with certain cloning methods...shuttle plasmid, verified, and linearized with the restriction enzyme PmeI . This construct is then transformed...plasmids are then verified for size and proper restriction digest patterns to determine that the transgene...two (BSL-2). They are highly immunogenic and can trigger a strong immune response to researchers if exposed...34354082 (Link opens in a new window) Rosewell, A., Vetrini, F., & Ng, P. (2011). Helper-dependent adenoviral...polyhedron with 20 faces, each being an equilateral triangle. Immunogenicity The ability of a molecule or substance...

  8. Gamma-Retroviral Vector Guide

    Type
    Guide
    ...with most standard cloning methods, including restriction enzyme, Gibson Assembly, or Gateway. Some transfer...transfer plasmids may have limited restriction sites or may only be compatible with certain cloning methods...the viral particles is removed and stored or centrifuged to concentrate. Crude or concentrated virus can...gamma-retroviral vector purification by high-speed centrifugation for CAR T cell manufacturing. Molecular Therapy...structural protein of the retroviral particle containing matrix, capsid, and nucleocapsid components. pol in trans...

  9. CRISPR Guide

    Type
    Guide
    ...activity with endogenous cell-type specific miRNAs to restrict CRISPR editing only to the target tissue. Browse... Acidaminococcus sp. ). The popularity can be attributed to Cas12a’s many benefits. Cas12a exhibits increased... quantification and can be used on lateral flow strips. Other Cas enzymes can be added to SHERLOCKv2 to...encyclopedia of Class 2 CRISPR systems with wiki entries describing enzyme activity, experimental considerations...924–927. PMID: 30377362 Casini, A., Olivieri, M., Petris, G., Montagna, C., Reginato, G., Maule, G., Lorenzin... (12), 770–788. PMID: 30323312 Yan, J., Oyler-Castrillo, P., Ravisankar, P., Ward, C. C., Levesque, S....152 (5), 1173–1183. PMID: 23452860 Savic, D., Partridge, E. C., Newberry, K. M., Smith, S. B., Meadows...

  10. Guide to Using Pooled Libraries

    Type
    Guide
    ... or identifying previously unknown genes that contribute to a phenotype. At their simplest level, pooled... the majority of the genome, while others are restricted to certain gene sets. In cDNA libraries, each...anti-cancer drug vemurafenib , resistance to Clostridium septicum alpha-toxin , and resistance to to the... Negative Screen Negative screens are a little trickier than positive screens. In a negative screen, the...

  11. Plan Your Experiment

    Type
    Guide
    ... containing the gRNA scaffold using standard restriction-ligation cloning. However, the exact cloning ...agarose gel to identify DNA fragments. PCR and restriction digest (for HDR repaired DSBs): For small nucleotide...nucleotide edits that introduce a novel restriction site. The region of interest is PCR amplified, digested...digested with the appropriate restriction enzyme and run on an agarose gel to identify DNA fragments. PCR...

  12. Optogenetics Guide

    Type
    Guide
    ...circuits. Optogenetic tools have also been used to trigger and study certain behavioral responses in model...hyperpolarization (inhibition) of the cell when triggered with yellow light, thus inhibiting function of... that hyperpolarizes (inhibits) the cell when triggered by green-yellow light. Arch Variants. Arch variants...its natural partner PpsR2 from Rhodopseudomonas palustris bacteria. Key feature is the sensitivity to 740...

  13. Lentiviral Vector Guide

    Type
    Guide
    ...with most standard cloning methods, including restriction enzyme, Gibson Assembly, or Gateway. Some transfer...transfer plasmids may have limited restriction sites or may only be compatible with certain cloning methods...containing the viral vectors is removed and stored or centrifuged to concentrate. You can use crude or concentrated...structural protein of the lentiviral particle containing matrix, capsid, and nucleocapsid components. pol in trans...

  14. Educational Resources

    Type
    Guide
    ...Learn about plasmids, sequencing, vectors, and restriction enzymes to assist with your molecular biology...Plasmid Cloning Creating Bacterial Glycerol Stocks Restriction Digests Guides Browse our in-depth instructional...

  15. Science Guides

    Type
    Guide
    ... the majority of the genome, while others are restricted to certain gene sets. Read More Viral Vectors...

  16. Sequencing Primers

    Type
    Guide
    ...provided for your reference. Addgene does not distribute primers. For sequencing plasmids in our repository...

  17. Chemogenetics Guide

    Type
    Guide
    ...Selective Modulation of cAMP Production in Striatopallidal Neurons. Neuropsychopharmacol. 38:854–862. ...

  18. AAV Production in HEK293 Cells

    Type
    Protocol
    ...8000 (PEG) + 0.5 M NaCl Cell lysis buffer (50 mM Tris HCl, 150 mM NaCl, 2 mM MgCl2) Reagent Preparation...Aliquot and store at 4 °C. Cell Lysis Buffer : 50 mM Tris HCl, 150 mM NaCl, 2 mM MgCl 2 Add the following ... bottle: 1836 mL deionized water + 100 mL of 1 M Tris HCl pH 8.5 + 60 mL of 5 M Sodium Chloride + 4 mL...-Cap), plasmid expressing your gene of interest Triton X-100 Benzonase/DNAse I (Millipore 71205-3) 40%...tilting the CS5 back and forth. Pro-Tip To help distribute the media amongst the five layers, tilt the CS5...container. Gently pour new media into the CS5. Distribute the media evenly between the layers. Return the...PBS and add to the harvested media and cells. Centrifuge at 3900 rpm for 20 min at 4 °C to pellet the ...

  19. Handling Plasmids from Addgene - Purifying Plasmid DNA

    Type
    Protocol
    ...solutions: Solution I - Resuspension Buffer 25 mM Tris-HCl (pH 8) 50 mM glucose 10 mM EDTA Store Solution...towel for 5-20 min. Resuspend dry DNA with TE (10 mM Tris-HCl pH 8, 0.1 mM EDTA). Pro-Tip DNA resuspension...Last Update: Feb. 8, 2018 Equipment Desktop microcentrifuge Desktop vortexer Vacuum (optional) Reagents... (low copy plasmids require larger cultures). Centrifuge the culture to pellet the bacteria before proceeding...entire overnight culture cannot fit into a single centrifuge tube, aliquot it into several tubes/bottles. ...solution. Pellet the proteins and genomic DNA by centrifugation, and remove the plasmid-containing supernatant...the stock culture to a 1.75 mL microfuge tube. Centrifuge in microfuge tube at 10,000 g for 30 sec. Pour...

  20. Plasmid Modification by Annealed Oligo Cloning (with Protocols)

    Type
    Protocol
    ... should be resuspended in annealing buffer (10mM Tris, pH7.5-8.0, 50mM NaCL, 1mM EDTA) and mixed in equimolar... Annealed Oligo Cloning You may also like... Restriction Digest of Plasmid DNA DNA Ligation Bacterial ...this tutorial we will discuss how to add new restriction sites to the MCS of an empty vector. However,...design a set of oligos containing your desired restriction sites and add them to your existing vector. It...cloned directly into the overhangs generated by restriction digest of existing sites in the original vector...waiting for your oligos to arrive, conduct a restriction digest of 1μg of vector with EcoRI and SalI. ...
Showing: 821 - 840 of 885 results