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We narrowed to 326 results for: cloning

Showing: 81 - 100 of 326 results

  1. Evolution of Lab Techniques

    Type
    Blog Post
    ...Restriction Cloning Get Tips on Analyzing DNA Sequencing Results Check out Plasmid Cloning by PCR Resources...applications range from confirming the results of cloning or knockout experiments to forensics and clinical...Labs still keep restriction enzymes on hand for cloning and analytical digests. A researcher can test the...sequencing, such as confirming the results of a cloning experiment, require only one or a few sequencing...

  2. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

    Type
    Blog Post
    ...References Geertsma ER (2013) FX Cloning: A Versatile High-Throughput Cloning System for Characterization ...efficiently subcloned into the phagemid pDX_init plasmid, using the fragment exchange (FX) cloning technique...pools from the phage display screens are subcloned (using FX cloning) into the expression vector pSb-Init,...for identification. All unique sybodies can be subcloned into pBXNPH3 or pBXNPHM3 expression plasmids for...

  3. Delivery Methods for Generating iPSCs

    Type
    Blog Post
    ...compared to other delivery methods. Retrovirus has a cloning capacity of ~8 kb, so either a single polycistronic...random integration creates variation between iPSC clones. Retroviruses are not considered safe for clinical...dividing and nondividing cells and have a larger cloning capacity (8-10 kb) than other retroviral vectors...cell, so there’s unavoidable heterogeneity between clones. This can complicate comparisons made between samples...between a vector and chromosomal DNA. PB has a cloning capacity of ~9-14 kb. This system is transfected...generating iPSCs by plasmid transfection where 2 of 11 clones tested had plasmid integration. Another challenge...for iPSCs Characterization Once generating iPSC clones, it’s important to determine how stem cell-like...

  4. Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs

    Type
    Blog Post
    ...Observing a bright signal for the first time after cloning an FP fusion with your favorite protein of interest...C-terminus [3]. You must therefore determine your cloning strategy and decide whether you will create an ... experimental needs. Equally important, before cloning you must consider whether and how well your FP ...peptide tagging to detect proteins expressed from cloned genes: deletion mapping functional domains of Drosophila...

  5. Sharing Your Lab Protocols: Using Apps to Save Time & Track Your Experiments

    Type
    Blog Post
    ...Recently I learned that Addgene’s pLKO.1 cloning protocol is viewed around 3,000 times each month. I ...Addgene website in December 2006, the pLKO.1-TRC cloning vector (deposited by David Root of the Broad Institute...has also cataloged a variety of useful molecular cloning protocols, including making LB agar plates, restriction...), and now Addgene. I just began uploading our cloning protocols to the site and will keep you updated...protocol listed under “Supplemental Document”: gRNA_Cloning Vector (deposited by George Church’s lab). Addgene...

  6. How to Design Your gRNA for CRISPR Genome Editing

    Type
    Blog Post
    ... and usually involves the need for single cell cloning and subsequent screening for successful edits. ...requires not one but two rounds of single cell cloning – as a control, one should revert the edit back...passenger variant that came along with the single cell clone (although this is rarely done). When targeting a...Veres et al., 2014). When working with single-cell clones, the authors note that “clonal heterogeneity may... CRISPR-Cas9 and TALEN Targeted Human Stem Cell Clones Detected by Whole-Genome Sequencing. Cell Stem ...

  7. Scientists Map the SARS-CoV-2-Human Interaction Network

    Type
    Blog Post
    ...flanked by EcoRI and BamHI restriction sites - so cloning projects should be fun and straight-forward. All...have an IRES-Puro marker to facilitate stable cell clone selection. Expression is driven by a fairly strong...study - so you might want to be careful with these clones (and help us figure out what is going on there!...

  8. Improved Plasmid Maps Powered by SnapGene

    Type
    Blog Post
    ...are commonly used for Golden Gate cloning or for CRISPR gRNA cloning. With this more functional display...display, you can start thinking about your next cloning experiment early on. Beyond these simple but powerful...

  9. Plasmids 101: Broad Host Range Plasmids

    Type
    Blog Post
    ...host range plasmids are more stable, can be easily cloned in different bacteria, and can be transferred to...inducibility (Toh et al. 2023), and additional cloning sites (Keen 1988).  Find broad host range recombinant...1988). Improved broad-host-range plasmids for DNA cloning in Gram-negative bacteria. Gene, 70(1), 191–197...R. (1988). Chapter 15—Broad-Host-Range Plasmid Cloning Vectors for Gram-Negative Bacteria. In R. L. Rodriguez...

  10. Adeno-associated Viruses (AAVs) for Genome Editing

    Type
    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...

  12. More Data for You: Find Articles Citing Addgene Plasmids

    Type
    Blog Post
    ... help save you from performing any unnecessary cloning - the list of "Associated Plasmids" from these ...associated with this "Citing Article" - such as cloning their gene of interest into a previously deposited...

  13. Antibodies 101: Polyclonal Antibodies

    Type
    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...

  14. New Tool for Lineage Tracing: The ClonTracer Library

    Type
    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...

  15. Pairing CombiGEM and CRISPR for Combinatorial Genetic Screening

    Type
    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...

  16. Antibodies 101: Choosing the Right Isotype

    Type
    Blog Post
    ...Monoclonal antibodies contain a single antibody clone and therefore a single isotype while polyclonal ...advantages. Since monoclonal antibodies contain a single clone they have higher lot to lot specificity and tend...Since polyclonal antibodies contain a variety of clones and thus the potential to bind many different sites...

  18. Pathways Over Time Plasmids Engage Students in Functional Genomics Research

    Type
    Blog Post
    ... strains are available. Then, staff or students clone similar sequences from a less-well studied organism...enable multiple student experiments We used TOPO cloning to insert the coding sequences for ten different...plasmid is controlled by the GAL1 promoter. The cloning procedure adds both V5 and His6 epitope tags to...

  19. Plasmids 101: Knockout/Knock-In Plasmids

    Type
    Blog Post
    ...to exons 1 and 3 of the targeted gene have been cloned into the vector on either side of an antibiotic...For this purpose, the sequence to be inserted is cloned into the vector between the homologous sequences...the one shown below, where the sequence of GFP is cloned together with the Neomycin resistance (NeoR) gene...
Showing: 81 - 100 of 326 results