We narrowed to 855 results for: NTS

...from their contents. You should be tracking both, you should easily be able to move contents from one container...can be useful in academic labs where postdocs, students, and lab mates are coming and going frequently...Addgene website, it sets off a chain reaction of events throughout all of Addgene - within the office, ...applied to containers and not the mushy, gooey contents of said containers, think carefully about how ...encoded numbers. Loosely couple containers and contents Right from the get-go be as clear as you can be...are being applied to the containers, not to the contents. Tightly coupling barcodes and containers might...such a bad idea, tightly coupling them to their contents might leave you sad and confused when a tricky...

...1-3 kb (approximately gene-sized) fragments so that these fragments can be cloned into E. coli and subsequent... Express random phage DNA fragments in E. coli Now that random DNA fragments have been cloned into E. ...return no candidates. Our strategy: Express random fragments of the phage genome and probe with live cells ...DNA frequently enough to result in a smear of fragments that get smaller in size as a function of digestion... method that results in the bulk of resultant fragments falling between 1 and 3 kb results in optimal ... causing issues associated with cloning large fragments. After fragmentation, repair sheared ends with...into chemically competent E. coli and analyze transformants for appropriately sized inserts. We consider...

...fixation/permeabilization reagents for intracellular targets. Since knockout experiments can be time-consuming...selectivity of an antibody for flow cytometry experiments. This will include a discussion of the Human ...approach that prioritizes more reproducible/renewable reagents, such as recombinant or hybridoma-derived monoclonal...cells that are amenable to genetic knockout, this represents a very robust approach to antibody validation...staining. This can include using: different blocking reagents; titration of antibody concentration (this can... need to study protein expression at multiple timepoints. When the target of interest is critical to cell...increase confidence that the labeling observed represents the expression level of the target of interest...

... easily access the documents. These tools not only allow users to access documents from anywhere, but ... papers on your desk, and to tackle some new experiments before the next R01 is due. But what about organizing...re frustrated and no closer to starting your experiments. Get organized now! Start by gathering that pile... color coded binders and tabs (for different experiments or projects) to make it easier to know where ...go Never miss a deadline again. Organize you experiments, keep track of upcoming grant deadlines, and ...from webpage clippings, photographs, and file attachments. Organize your notes into notebooks, create checklists...required to change the status quo. But with recent improvements to ELN and online lab management software, there...

...selectable DNA fragments, such as antibiotic resistance genes, as well as non-selectable DNA fragments, such as... as gene replacements and tags.  The typical frequency of recombinants is 1 positive clone out of 104 ...Enhanced levels of λ Red-mediated recombinants in mismatch repair mutants. Proceedings of the National Academy...homology to the target site for recombination. Key components of the Lambda Red recombineering system The lambda... lambda red recombineering system has three components (Figure 1): 1) Exo, 2) Beta, and 3) Gam. All three...modification with an ssDNA substrate.  Gam: Gam prevents both the endogenous RecBCD and SbcCD nucleases...an ssDNA oligo substrate.   Figure 1: Components of the Lambda Red Recombineering system. See ...

...cell to perform multiple recombination events, all linear fragments need to get into the same cell. For ...3), producing multiple fragments with specific homologous ends. These fragments are assembled in vivo ...multiple recombination events to assemble the final plasmid. At least 5 fragments can be correctly assembled...Figure 1). As this approach requires no special reagents or purification kits, it is low cost and can be...protocol:  Fragment generation - Any linear DNA fragments can be assembled in vivo as long as they have ...PCR is the primary method to produce the linear fragments, as desired modifications and homologous regions...positive colonies. Transformation - the linear fragments are transformed into E. coli and plasmid assembly...

...essential components of both pathways have been shown to be efficacious for increasing knock-in events (Yang...mutations and gene editing Targeted genome editing events fall into two general classes: knock-in and knock-out...frameshift mutation. The majority of cellular repair events will generate these types of mutations, making ...DNA repair mechanism in general. On average, it accounts for less than 10% of DNA repair, with this rate...considerations for designing donor DNA for successful HDR events. CRISPR cut site location Position your CRISPR...disrupting mutations in donor DNA   Promoting HDR events over other edits Like I said earlier, HDR is not... to exist as a repair template. To maximize HDR events, ensure cells are actively cycling through the ...

...of the vector components are HIV-based? A7: No single plasmid contains all the components necessary to ...The components are divided as follows: Transfer vectors contain minimal cis-acting HIV components: LTRs...specific experiments being performed and you should understand the guidelines for using such reagents as outlined... produced by co-transfecting three essential ingredients: the lentiviral packaging vector, a transfer ...Second-generation vectors are likely sufficient for most experiments; however, the 3rd-generation packaging systems...the 3'LTR in the transfer plasmid. The deletion prevents full-length viral RNA from being produced in target...LTRs, PPT, RRE, and psi packaging signal. Viral components typically total <1.5kb, which is rarely more ...

...NGCG 8  Tips for designing base editing experiments in plants Choose a base editing platform depending...engineering through adenine base editing in plants." Nature plants (2018): 1. PubMed PMID: 29867128. 11. Li...Plant base editor platforms and their molecular components. Adenine base editors for plant We developed... could be used to generate stable genome edited plants via Agrobacterium-mediated transformation. We also...). Type Nickase Cas9 (D10A) Deaminase PAM requirements Reference CBE SpCas9 rAPOBEC1 NGG 6 SpCas9...transient assay. For stable transformation, regenerated plants may be subjected to genotyping. If successful base... Yuan, et al. "Efficient C-to-T base editing in plants using a fusion of nCas9 and human APOBEC3A." Nature...

...result presents an opportunity for troubleshooting and optimization of the transfection reagents and process...There are many, many different types of experiments carried out by scientists every day. Although the... controls that can be used for plasmid-based experiments. What are control plasmids? Generally, control...interpretation of the results. Typically, plasmid-based experiments employ transfection, negative, positive, and ...control allows you to examine if the transfection reagents or the transfection process itself has any cytotoxic... on gene expression in your target cells. In experiments employing gene targeting or genome editing technologies...results are reproducible from properly controlled experiments. The observed phenotype associated with the independent...