We narrowed to 324 results for: cloning

...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 ...

...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...

...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...

...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...

...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...

...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...

... 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 ...

...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...

...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...

...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 ...

... 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...

... 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...

...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...

...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...

...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...

...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...

...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...