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...related LCR technique uses two sets of probes to achieve exponential amplification. What does fidelity mean...substrates. Thus, high fidelity ligation can be achieved through a combination of the intrinsic selectivity...four upstream probes and four down-stream probes, each differing only in the base at the ligation junction...represented. Sixteen separate pools were prepared, each with a different complement strand covering all ...the template. The probes were designed such that each possible pairing resulted in a product of a different...

... on genome sequencing. Used in concert, these approaches can provide the researcher with a reasonable ...(Fu et al. 2013; Pattanayak et al. 2013). This approach misses larger mismatches (e.g. six nucleotides... may be to integrate both biased and unbiased approaches. As suggested in a review by Tycko et al., 2016...a highly accurate manner. Using both of these approaches may not be necessary for every CRISPR experiment...Mark J., et al. "Evaluation of TCR gene editing achieved by TALENs, CRISPR/Cas9, and megaTAL nucleases....

...found yourself staring at a browser full of tabs, each open to a different antibody option. Or you may ... options in the Antibody Data Hub. Data reports  Each antibody submission is used to create a data report...Curating the data reports Just like our AAV Data Hub, each data submission we receive is checked by Addgene's... Rachel Leeson...

...a plasmid or viral origin. The type of cloning approach you choose ultimately will influence the quantity... for the extraction. Restriction enzyme based approaches can require several hundreds of nanograms or ...assembly or Golden gate assembly use a more direct approach, allowing scientists to clone from a smaller amount...will likely require micrograms of plasmid DNA for each reaction. As a consequence, the preparations for...

...light used to activate the first PA-FP does not photobleach the second, and vice versa. Finally, as every...contrast, PA-FPs offer a reduced potential for photobleaching and phototoxicity that standard FPs might induce... PA-FPs is photoactivated, imaged, and then photobleached. Additional PA-FP molecules are then activated...still do not emit the large number of photons achievable with photoswitchable dyes. Correlative light ...297.5588 (2002): 1873-1877. PubMed PMID:12228718. 2.Subach, Fedor V., et al. "Photoactivatable mCherry for...

...facts. Because, similar to scientists, the legacy of each government and generation is in the veracity of ...the lab, she dabbles in science policy, public outreach, and restorative justice.  Follow her on twitter...where these issues hit hardest. According to the Guttmacher Institute, most women of reproductive age live...galilei-dialogues-concerning-two-new-sciences). This is the approach policy makers take in abusing climate science ...tirelessly to get the publications, grant funding, and teaching experience that it takes to complete your resume...

...consensus lentiviral expression vector and attached a DNA barcode to each to make the library usable for pooled...proliferation, survival, differentiation, and apoptosis. For each of these 17 pathways, they identified a set of one...researchers didn’t have to discover new ways to activate each of these pathways. Rather, they relied on published...

...method that uses iterative staining and chemical bleaching to allow for high-resolution imaging without degrading...IBEX: An iterative immunolabeling and chemical bleaching method for high-content imaging of diverse tissues...). IBEX: A versatile multiplex optical imaging approach for deep phenotyping and spatial analysis of cells... Rachel Leeson...

...2014). Table 1. Summary of Cas9 domains, the lobe each domain belongs to, and their roles. Lobe Domain...crRNA:tracrRNA duplex — Cas9’s two lobes move toward each other into the “closed” conformation shown in Figure...test DNA bases that are already snugly paired to each other? At this point, the protein can switch between...guide RNA (Cofsky et al., 2022; Osuka et al., 2018). Each matching DNA:RNA pair gives just enough stability...own advantages—and blind spots—to these questions. Each new structural insight also spurs the next generation...

..., I reached out to Amy with exciting news. I managed to secure a virtual summer program, teaching me the.... A capstone project is a major project given to each intern to complete and present on.    Figure...report shared my work by breaking down the process of each day, my background, observations, etc. On my final...

...users have recently begun to develop CRISPR-based approaches for studying gene function in these systems. ...use in a fetal-lung derived organoid system, the approach is intended to be amenable to organoids derived...tissue-types. By bringing together CRISPR-based approaches and organoid models, toolkits like this one provide...trigger seed germination of parasitic plants, this approach was also used to build and validate a ratiometric...

...University and entered a new role here at Addgene as an Outreach Scientist. I used to spend my days (and often ...This is particularly true in my position as an Outreach Scientist where I spend a large amount of my time...communicator that got me here! Thus far being an Outreach Scientist at Addgene has been awesome. The process...researchers gain doing PhDs (i.e. technical writing, teaching, presenting, analytical thinking, time management...

...another method Transfection and transduction seem to achieve similar outcomes, so why and when should you opt...Second, what is the biological outcome you want to achieve? For example, do you want the nucleic acid cargo...work is that you can swap out your virus type to achieve different biological outcomes. For example, some...targeting of that virus, which can’t be easily achieved with alternative methods. Ready, set, transduce...

...disease front. Viruses survive by hijacking the machinery of the cells they infect. Once they enter a cell...delivered to the cell. The cell then uses its own machinery to replicate the viral genome and proteins. This... growing number of Viral Vectors 101 posts about each type, so be sure to check our blog, or our Viral... Rachel Leeson...

...surface. Their toolkit includes a set of plasmids each containing an anti-GFP nanobody fused with a different...genomic locus: Caspex  Article contributed by Susanna Bachle  Listen to the Caspex podcast segment Steven Carr...Scientist in the group, developed a CRISPR-based approach to detect proteins that associate with specific...Sontheimer and his group from the University of Massachusetts Medical School have developed a group of plasmids...

...assays, in vitro transduction, and silver stain approaches we show that there is no difference in purity...so excited? For starters, we believe that this approach can be adapted by vector cores and laboratories...investigated 3 serotypes it’s possible that this approach will be beneficial for a host of other AAV serotypes...serotypes; it will be exciting to test this approach with different capsids. Most importantly, because ...

...a smaller RNA tag, are less prone to photobleaching, and achieve better signal in vivo as compared to ...the second containing the growth promoting domain. Each half was fused with one of the LOVTRAP domains. ...dCas9 fused to cytidine deaminases like APOBEC. To achieve this they created a lentiviral vector system that...

...chromosomal DNA, but they still rely on the host machinery to make additional copies. ori sequences are generally...lower temperatures and allows the replication machinery room to come in and get busy making copies.   ...incompatible because they will compete for the same machinery, creating an unstable and unpredictable environment...higher copy numbers since the bacteria have just reached stationary phase, but the cells have not started...

... unexplored. Using new single-cell sequencing approaches, the lab was able to eliminate the need for axenic...YNP, it took some time to map a route that would reach all of the remote areas, including one hot spring...poles to collect water samples from a distance. At each collection site, we took GPS coordinates, water ...was separated into smaller samples of 5-10 cells each for sequencing, allowing for lower complexity with...

...Below, you will find some easy talking points for broaching this subject with your institution’s tech transfer...Agreements. By default, your TTO must sign an agreement each time a material is being requested or deposited....Reception and Master Deposit Agreements, your TTO signs each agreement just once, covering nearly all future ...University of California, Irvine University of Massachusetts, Worcester  University of Nebraska Medical Center...