...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...

... 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 ...

...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!...

...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...

...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...

...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...

... 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...

...seen with things like DNA synthesis, Golden Gate cloning, and CRISPR. The yeast synthetic genome project...Nonprofit BioBuilder Plasmids 101: Golden Gate Cloning Learn About Working at a SynBio Startup in Our ...

...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...

... 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...

...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...

...are getting better and better at identifying and cloning the genes for antibody expression. If starting ...through DNA sequencing of the line and subsequent cloning of the antibody chains (Crosnier et al., 2010; ...proteins. After the antibody of interest has been cloned into an expression plasmid, the plasmid can be ...

... 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...

...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...

...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...

...plasmids uses the same restriction enzymes for cloning (see schematic below), enabling a researcher to...tags Resources on Addgene.org Find Additional Cloning Kits ...

...terminus the tag will end up on after successful cloning. Cloning information: This section provides useful ...plasmid's structural features and see if I can easily clone my gene into it? Your best bet is the Plasmid ...

...were used in this study.  The zCas9i cloning kit is a plant cloning kit that contains a series of vectors...CRISPR-based lineage tracing tool captures rare clones that represent 0.001% of the population.  Because...

...present in these mice, you can clone the PCR products into a pBluescript cloning vector and sequence the resulting...region into a pCAG-EGXXFP plasmid using standard cloning techniques (The XX indicates where your target ...

...without a kit Read our blog posts about plasmid cloning Browse our Plasmids 101 blog posts Resources on...Molecular Biology Reference Find protocols on plasmid cloning Find plasmids for your research ...