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  9. Using AAV for Neuronal Tracing

    Type
    Blog Post
    ... natural RABV or HSV can). AAV does not have a membrane envelope like lentivirus, but instead is contained...

  10. Cloning

    Type
    Guide
    ...exonuclease. Gibson Assembly (Image from Plasmid 101: Gibson Assembly ) Back to Top Type IIS Assembly (Golden Gate...on TOPO cloning. Back to Top Gibson Assembly (Isothermal Assembly Reaction) Isothermal cloning, more commonly... commonly known as Gibson assembly ( protocol ), takes advantage of the properties of 3 common molecular...DNA ligase fuses DNA strands together. In Gibson assembly DNA fragments with 20-40 base pair homology at...Gibson cloning is that it allows for the simple assembly of multiple fragments of DNA in the chosen orientation...into. A drawback of this system is that Gibson assembly works best when combining DNA fragments over 200...custom overhangs that can then be efficiently assembled together. The advantage of type IIS systems are...

  11. Optogenetics Guide

    Type
    Guide
    ...attaching only one member of the Zdk/LOV2 pair to the target protein, and the other to the membrane where the ...trafficking signals to localize opsins to the cell membrane. Microbial opsins, such as those described below... at physiological pH and trafficks well to the membrane. Excitatory (Depolarizing) ChR Variants. The optogenetic...from Kir2.1 and ER export signal provide improved membrane targeting 589 Archaerhodopsins Arch Light-activated...experimental contexts, including protein activation, membrane localization, and transcriptional activation. ...reversibly sequestering targets into large, optically assembled protein architectures in living mammalian cells...light-inducible dimers; Bacterial SsrA peptide is embedded in the LOV2 domain (iLID plasmid), while its natural...

  12. Antibody Guide

    Type
    Guide
    ...Transfer proteins from the gel to a membrane. Incubate the membrane with an unconjugated primary antibody...block the membrane to remove excess antibody and prevent unwanted binding. Incubate the membrane with a ...Wash and block the membrane. Activate the HRP with a substrate kit. Image the membrane using either X-ray...tears, saliva, mucus) where it protects mucosal membranes Dimer with a valency of 4 IgA1; IgA2 IgD Coexpressed...short peptide linkers. Diabodies can be used for assembling protein nanostructures. Figure 2: An antibody...recognizes your primary antibody source species. Remember that recombinant antibodies will also have a source...

  13. Adeno-associated virus (AAV) Guide

    Type
    Guide
    ...and Cap regions necessary for the production and assembly of viral capsids. Helper plasmid — containing ... methods, including restriction enzyme, Gibson Assembly, or Gateway. Some transfer plasmids may have limited...infect cells and replicate autonomously. Human embryonic kidney 293T (HEK293T) cells are the most common...rAAV vector (∼5%). Gene fragments can also be reassembled via homologous recombination, when both sequences... infection risks occur with contact to mucous membranes or broken skin. Needle sticks and ripped gloves...

  14. Addgene's eBook Collection

    Type
    Guide
    ...Select Plasmids 101 September 2023 | Fourth Edition About Plasmids 101 September 2023 | Fourth ...Science Career Guide November 2018 | First Edition About Science Career Guide November 2018 | First...

  15. Adenovirus Guide

    Type
    Guide
    ...primarily encode for structural proteins needed to assemble icosahedral capsids and build new virions. Figure...by the packaging cell line, normally the human embryonic kidney 293 (HEK293) cell line. There is a rare... methods, including restriction enzyme, Gibson Assembly, or Gateway. Some transfer plasmids may have limited... infection risks occur with contact to mucous membranes or broken skin. The risk of exposure and reaction...

  16. Gamma-Retroviral Vector Guide

    Type
    Guide
    ... methods, including restriction enzyme, Gibson Assembly, or Gateway. Some transfer plasmids may have limited...Gamma-retroviral vectors can be packaged directly in human embryonic kidney 293T (HEK293T) cells (Figure 3A), similar... infection risks occur with contact to mucous membranes or broken skin. Needle sticks and ripped gloves...

  17. Lentiviral Vector Guide

    Type
    Guide
    ... methods, including restriction enzyme, Gibson Assembly, or Gateway. Some transfer plasmids may have limited...requires a production cell line, typically human embryonic kidney 293T (HEK293T) cells. Standard HEK293T ... infection risks occur with contact to mucous membranes or broken skin. Needle sticks and ripped gloves...

  18. Molecular Biology Reference

    Type
    Guide
    ...nucleotide is present at the end of each fragment and assembles that information into the DNA sequence. Sanger...reactions on different pieces of DNA to be able to assemble the whole puzzle. That’s where Next Generation...sequenced templates are then aligned to each other to assemble the entire sequence or puzzle. It is important...

  19. CRISPR Guide

    Type
    Guide
    ...against these foreign nucleic acids. Acr family members inhibit CRISPR by a variety of mechanisms, including...Mediated Genetic Screening in Mice with Haploid Embryonic Stem Cells Carrying a Guide RNA Library. Cell ...

  20. Plan Your Experiment

    Type
    Guide
    ...off-target effects Can be used to generate transgenic embryos Cas9-gRNA ribonucleoprotein (RNP) complexes Purified...

  21. Chemogenetics Guide

    Type
    Guide
    ...cell surface receptors, GPCRs are seven-pass transmembrane proteins that bind a specific ligand. Ligand...
Showing: 601 - 621 of 621 results