We narrowed to 457 results for: nsf

...false positive colonies. Transformation - the linear fragments are transformed into E. coli and plasmid...assembly of a circular plasmid in vitro, before transforming it into E. coli for propagation. However, while...using a simple two-step, 2 hr protocol prior to transformation (Figure 1). As this approach requires no special... encode homology arms to allow immediate co-transformation with linearized vector backbone. DpnI digestion...failed so far. What is most important is the transformation efficiency of the strain: when asking the cell...

...of three plasmids: the packaging, envelope, and transfer plasmids. The packaging plasmid carried a mutated...of cells the vector could infect. Finally, the transfer plasmid encoded the desired transgene flanked ...packaging and host genome integration. After co-transfection of these plasmids, 293T cells released transgene-containing...-competent virus via recombination between the transfer and packaging plasmids does exist. To further ...work with primary cells or other difficult-to-transfect cells. Lentiviral barcoding libraries have been... 18;127(7):839-48. PubMed PMID: 26758916. 7. Transfusion independence and HMGA2 activation after gene ...

...opportunity to discover and evolve your transferable skills. Transferable skills are the non-technical skills...formulate hypotheses and conduct experiments. You transfer liquids between tubes, look through microscopes...production, I didn’t really start to recognize my transferable (or “soft”) skills until I was out of the lab...lab to think about these--get started with our Transferable Skills Guide.     Over the last decade, my... blog Visit our careers topic page Read our transferable skills guide Read blog posts about career readiness...

...72 hours post-transformation/transfection in a transient assay. For stable transformation, regenerated ...deaminases (ecTadA), fused it with dCas9, and transformed it into E. coli harboring the defective CamR ...genome edited plants via Agrobacterium-mediated transformation. We also constructed smaller vectors suitable...suitable for transient assays using protoplast transformation. Cytosine base editors for plant There are several...

...editor consisting of dCas9 fused to a DNA methyltransferase — that can drive lasting transcriptional silencing...Histone tail for Autoinhibition Release of Methyltransferase (CHARM), with several advantages over the ...  Rather than overexpressing an exogenous methyltransferase for epigenetic silencing, which can be toxic...tail to recruit and stimulate the endogenous methyltransferase. The strategy is generalizable to a variety...D3L recruits the endogenous, autoinhibited methyltransferase DNMT3A to the genomic location specified by...DNA-binding protein. The ADD domain of the methyltransferase binds to H3K4me0 on D3L, releasing the autoinhibition...

...certain modifications to DNA bases.  Ligate and transform your resultant plasmid libraries into chemically...chemically competent E. coli and analyze transformants for appropriately sized inserts. We consider a library...culture, dilute and plate the complete library of transformants for single colonies (this usually requires 10...10-20 plates) (see box below). This initial transformation should be performed using a highly competent...extract plasmid DNA directly from the colonies. Transform the resultant plasmid DNA into E. coli BL21 (DE3... protein production), then dilute and plate transformants for isolated colonies (150-200 per plate) across...

...we have found that transfecting with PBx mRNA can boost efficiency over transfecting with plasmids in some...from a licensed distributor) allow you to quickly transfer a DNA sequence from the transposon vector to one...traditional method of making stable cell lines is to transfect, select, and isolate clones. This procedure is...integration efficiencies that are accomplished with transfections and are therefore virus-free. This is achieved...fully intact, while, with a standard DNA plasmid transfection, a double-stranded break may occur in your cargo...

...Differentiation Factors Find plasmids used to transform iPSCs and fibroblasts into neurons and more. Convert...

...worked closely with the legal partners and Tech Transfer Offices at universities to make COVID-19 research-related...United States, and many universities and Technology Transfer Offices were, at least, temporarily closing, these...scientists, we’ve worked with many scientists, tech transfer offices, research foundations, and funding initiatives...them available to industry, please reach out at techtransfer@addgene.org and we will initiate a conversation...conversation with you and your technology transfer office to discuss the possibilities for expanded sharing. ...

...Interest (GOI), and transfect it into your favorite cell line. In parallel, you transfect cells with another...express your protein of interest endogenously and transfect them with a plasmid encoding your protein of interest...expect to see evidence of antibody binding in the transfected cells but not in control cells. But (there is...results show a lovely, strong band from the cells transfected with your protein of interest and show just the... the faintest band from the cells transfected with the other plasmid. Your antibody works! Multiple (Independent...

...Resonance Energy Transfer (BRET). BRET is similar to Förster Resonance Energy Transfer (FRET), in which...fluorophore. The difference for BRET is that the energy transfer is between a luminescent donor and a fluorescent...dissociation by bioluminescence resonance energy transfer 2, or BRET2. With binding of the drug, the alpha...transducers. To use the TRUPATH system, cells are transfected with plasmids expressing the GPCR of interest...

...luminescent) worlds Forster Resonance Energy Transfer (FRET) is the transfer of energy from an excited donor fluorophore...Resonance Energy Transfer (BRET) works the same way, except that energy is transferred from a luminescence... fluorescent protein close enough, energy is transferred by BRET and the fluorescent protein emits light...

...strong enough to hold a plasmid together through transformation, so the cloned vector must be ligated. But ...nicks easily repairable by the bacteria after transformation. LIC is a reliable cloning method, but it is...recombination intermediate to bacteria, so it can be transformed as-is into cells. The cells then get to work ...is long enough to hold it together, it can be transformed into bacteria, which will efficiently repair ... annealed. The recombination intermediate is transformed into bacteria, which repair the plasmid. Created...

...useful for lineage tracing, evaluating infection/transfection efficiency, selecting desired clones, and making...

...Tethering dCas9 to the histone acetyltransferase p300 or the DNA methyltransferase DNMT3A allows researchers...demethylation?  Dividing cells depend on DNA methyltransferase 1 (DNMT1) to copy methylation marks from parental... al (2018). Genome-wide tracking of dCas9-methyltransferase footprints. Nat Commun 9, 597. https://doi.org...promoters by targeted steric blockage of DNA methyltransferase with CRISPR/dCas9. Nat Commun 12, 5711. https...

... Options for CRISPR delivery include plasmid transfection, RNP electroporation, and viral transduction... non-replicative. Nanoblades are produced by transfecting the following four components into 293T cells...lower off-target effects when compared to DNA transfection of gRNA and Cas9 expression plasmids. These ...having lower off-target edits compared to DNA transfection.  Homology directed repair (HDR) with Nanoblades...can then fuse with the target cell membrane, and transfer their CRISPR cargo into the zygote. As a proof-of-principle...

...easy — our tech transfer team will communicate with your organization’s technology transfer office or other... Send plasmid information to Addgene Material Transfer Agreement (MTA) approval  Send plasmids to Addgene...require that you send your samples to Addgene pre-transformed in this strain. Hazardous If the plasmid...samples to Addgene and contact your technology transfer office about the MTA. Pro Tip! Download a new...

...expression vector that is then transformed into a bacterial cell. Transformed cells propagate, are induced... strategy for protein expression consists of transfecting cells with your DNA template of choice and allowing...SV40, CMV, and RSV) for robust expression post-transfection. Mammalian systems can express proteins both... Both methods produce high protein yields if transfection is successful. Some mammalian systems also ...

...used by respondents (transfection method, amount of each component per transfection etc.). Take Pooran'...an ssDNA guide. Therefore, researchers must co-transfect cells with 5’-P-ssDNA guides and an NgAgo expression...delivery methods. Researchers can conveniently transfect/inject cells with in vitro preassembled Cas9/gRNA...researchers will have to carry out repeated transfections with the ssDNA guide, although many may find...

...determined by the different hosts that a plasmid can transfer to, replicate in, and persist in. Some plasmids...easily cloned in different bacteria, and can be transferred to various experimental hosts (Jain and Srivastava...scientific community, as they allow for simplified transformation processes, and can be used by scientists across...conjugative and carries the genes necessary for transfer initiation between bacterial hosts. Also, self-transmissible...Plasmids 101: What is a Plasmid?  Plasmids 101: Transformation, Transduction, Bacterial Conjugation, and Tranfection...