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We narrowed to 321 results for: cloning

Showing: 121 - 140 of 321 results
  1. Pairing CombiGEM and CRISPR for Combinatorial Genetic Screening

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    Blog Post
    ...synthesis using the format indicated below and pool-clone them into the pAWp28 storage vector (Figure 1): ...sequence and its 8-bp barcode, respectively. Once cloned into the storage vector, the pooled, barcoded gRNA... lentiviral vector backbone pAWp12 (Figure 2). Cloning from the pAWp28 storage vector to the pAWp12 lentiviral...
  2. Antibodies 101: Epitope Tags

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    Blog Post
    ...knock-in at a genomic locus or more commonly via cloning of recombinant proteins. Why use a tag? If you...developed in 1985 and is still the most prevalent clone used today! Goat, chicken, mouse, or rabbit? Addgene...
  3. Plasmids 101: Shuttle Vectors

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    Blog Post
    ...complex eukaryotic cells, but would prefer to grow, clone, and manipulate their vectors in fast-growing bacteria...example, if you are using E. coli simply for easy cloning and quick DNA production, you probably don’t need...called transfer vectors. The gene of interest is cloned into a shuttle vector for transfection into producer...
  4. High Complexity Golden Gate Assembly with NEB’s DAD

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    Blog Post
    ...If you’ve ever used Golden Gate Assembly for cloning, you might be familiar with the rules of thumb for...complexity allowable by a favored, fairly easy cloning technique (which may be of particular interest ...
  5. CRISPR 101: Off-Target Effects

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    Blog Post
    ...cell clones to use for multiple downstream experiments. You only successfully generate 1 null clone and...in that single clone is low, but since all downstream experiments will rely on that clone not having any... characterization of, isolating 2 or 3 distinct clones to perform key experiments on is a good way to ...
  6. Adeno-associated Viruses (AAVs) for Genome Editing

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    Blog Post
    ...synthesis company such as IDT or Genscript, and cloning them sequentially into the chosen acceptor vector...cells. After G418 selection, individual gene edited clones are identified by PCR. Then, if necessary for subsequent...knockin efficiencies of 1-40% with G418-selected clones. However, there are important disadvantages to ...AAV-based approach requires G418 selection of infected clones, making it unsuitable for most in vivo applications...
  7. Live and Let Dye: Self-Labeling Protein Tags

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    Blog Post
    ...ligands for different experiments without having to clone a whole new construct for every application. Plus...your labels easy! Instead of taking the time to clone a new construct for every new-and-improved fluorescent...able to get a plasmid ready-to-use without any cloning at all! As mentioned above, several variants of...collection to find self-labeling fusion proteins or cloning backbones to study your gene of interest with these...
  8. 15 Hot Plasmids from 2017

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    Blog Post
    ...that have standard cloning sites or gateway compatible cloning sites for easy cloning of your genes of ... and repair templates of choice though modular cloning steps. The Voytas lab plans to update the toolkit...
  9. Plasmids 101: E. coli Strains for Protein Expression

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    Blog Post
    ...of E. coli for DNA propagation. While great for cloning purposes, these E. coli strains are not usually... (DE3)** Rifampicin RecA-deficient; allows for cloning and expression in same strain F- recA1 hsdR(rK12...
  10. Fluorescent Proteins 101: Luciferases

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    Blog Post
    ...luciferase was the first bioluminescent reporter to be cloned and remains popular, other luciferases have been...gene regulation Since firefly luciferase was first cloned in 1985, luciferase assays have become a gold standard...— on gene expression. The regulatory element is cloned upstream of the luciferase gene, and expression...
  11. Antibodies 101: Polyclonal Antibodies

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    Blog Post
    ... the same isotype. But together, all the B cell clones in the immune system make different isotypes of... starts with selecting just one of these B cell clones for further antibody production, polyclonal antibodies...antibody increases the likelihood that one of the clones will still recognize your protein. For the same...
  12. New Tool for Lineage Tracing: The ClonTracer Library

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    Blog Post
    ...limit of roughly 0.1%. Thus, rare, pre-existing clones that are present at less than 0.1% of the total...only one barcode. The abundance of each barcoded clone can then be monitored over time by sequencing the...then corresponds to the abundance of a specific clone; lineage tracing is performed via barcode sequencing...
  13. CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More

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    ...template, using Gibson Assembly or restriction enzyme cloning. Streptomyces bacteria are more recombinogenic ...inducer, in this case anhydro-tetracycline (aTc). By cloning in two tandem copies of the gRNA cassette, one ...and the gRNA are Tet-inducible, and a gRNA can be cloned easily into pRH2521 using BbsI. This system results...syringae. These vectors contain BsaI sites for easy cloning of a gRNA of interest. Cytidine and adenine base...
Showing: 121 - 140 of 321 results