...plasmids. Want a retroviral plasmid with your gene of interest? Search our database for retroviral transfer plasmids...

...promoter or regulatory regions of your gene of interest. If the plasmid that you choose does not also ...

...20-basepair complementary region to any gene of interest. Co-expression of dCas9 and sgRNA can efficiently...

...easily visualize products from an AgeI/EcoRI restriction digest) David Root 10879 pLKO.1 ‐ TRC control...

...can be used as empty backbones for your gene of interest for expression in plants. You can search the table...

...genome are every 500 bp. TAL effectors have some restrictions (for example, the target must start with a T...

...instead encode for a specific gene or other research interest. To produce viruses with alternate (non-virus-...

... novel AAV vector targets ON bipolar cells and restores visual function in a mouse model of complete congenital...

...cell lines. Features include (i) Esp3I (BsmBI) restriction sites for full compatibility with the Golden ...

...reads from PCR amplicons that cover your locus of interest and correspond to edited and non-edited cell populations...

...template, you could then: Confirm your HR by doing Restriction Fragment Length Polymorphism (RFLP) ( see Figure...

...method, mouse strain, target cell type or tissue of interest, and model organism under study. *Many engineered...

...including ITRs), though there is a great deal of interest and effort directed toward expanding this capacity...

...very low or nonexistent. Scientists continue to wrestle with the possibility of postnatal and germline ...

...of R, is capped, and proceeds through U5 and the rest of the provirus. 3' LTR in cis Terminates transcription...plasmid encoding a transgene, sgRNA, or shRNA of interest: The transgene sequence is flanked by long terminal...

...exhaustive. If you do not see a particular gene of interest, please search our full plasmid collection using...

...of R, is capped, and proceeds through U5 and the rest of the provirus. Third generation vectors use a ...containing only one restriction site? If a lentiviral transfer plasmid contains a single restriction site, one ...subcloning or appending compatible restriction sites onto the insert of interest using PCR. The process of subcloning... basically shuffling restriction sites between vectors until the gene of interest is flanked by sites ...consuming and easier to simply add restriction sites onto the insert of interest using PCR. This is accomplished...using primers that contain the restriction sites needed. Functional restriction sites must be a certain number...Lentiviral transfer plasmid encoding your insert of interest. The transgene sequence is flanked by long terminal...

...a sequence recognized by the restriction enzyme MstII. When a restriction digest with MstII and southern...than real-time PCR (4). Restriction mapping and modern sequencing Restriction mapping informs on DNA sequence...different restriction enzyme digests. Incubating a DNA sample of unknown sequence with restriction enzymes...sequence of interest to create a physical map of the DNA. Starting in the 1970s, restriction mapping was...has changed, restriction enzymes are certainly not dead yet! Labs still keep restriction enzymes on hand...three historical techniques: southern blotting, restriction mapping, and sequencing gels, as well as their... (2). Southern blotting, in conjunction with restriction digest analysis, is used to identify the presence...

...Method C: Restriction cloning of a restriction enzyme fragment containing the DNA of interest and a attL-entry...scientists are no longer limited to traditional restriction enzyme cloning. Instead, you can choose a molecular...vectors, one can use Gateway to clone a gene of interest into a variety of expression systems. Keep reading...generates the entry clone containing the DNA of interest flanked by attL sites. As a byproduct of the reaction... a result, an expression clone with the DNA of interest flanked by attB sites is generated. As in the ...recombination, and it is exchanged with the gene of interest during the BP or LR reactions. Since the CcdB ...Figure 5: Method C to create an entry clone: Restriction cloning the inset into an attL-containing entry...

...the mutation of interest. Gel electrophoresis diagrams of the anticipated restriction analysis results... by Restriction Fragment Length Polymorphism (RFLP) analysis (right). Introduction of restriction site...conventional cloning (to introduce or remove restriction sites), in mapping of regulatory elements (to...the plasmid template (blue), and introduce a restriction site “A*” (in addition to the “A” site already...in the parent vector). Also shown are the “B” restriction sites, which flank the targeted region. Upon ...amplification, the template is removed by DpnI restriction digestion leaving behind only the mutagenized...primers to successfully anneal to the plasmid of interest during the PCR reaction. Ideally, your primers...