We narrowed to 798 results for: REP

...shared data, on our values of transparency, reproducibility, and community-building. The transparency begins...to better plan your experiments. To promote reproducibility, Addgene will have unique Research Resource...that recombinant antibodies are excellent for reproducibility because they do not share hybridomas’ genomic...drift or limited material from polyclonal animal preparations (Bradbury et al., 2015). All of Addgene’s antibodies...infrastructure, from our educational materials to our repository of open samples. We are, and have always been...2016) RRIDs: A Simple Step toward Improving Reproducibility through Rigor and Transparency of Experimental....2016.04.030    Bradbury A, Plückthun A (2015) Reproducibility: Standardize antibodies used in research. Nature...

...channel. And with no more ado, the questions!  AAV Prep and Technique Q1: Is there any way to deliver a ...you think it is possible to make an efficient AAV prep for a 4.2 kB insert with the CMV promoter using ...We have found this method to be reliable and reproducible. However, we do not use amplicons in ITR as ... in sample treatment across methods can affect reported titer by either method so to ensure equal animal...AAV8. Very recently an engineered AAV5 capsid was reported for specific and potent liver transduction. While...concentrated to ~1x1014 vg/ml.. As evident in our repository, this application is an outlier for PHP.eB (1x1011...enhanced CNS tropism to other serotypes, but reproducible success remains elusive.  There has also been...

... ready-made AAV and Lentiviral preps. This new Viral Service represents Addgene’s largest new initiative...16 labs currently sharing viral preps) through the Addgene repository. Without these scientists and the...institutions to distribute viral preps derived from plasmids in the repository. The Outreach Team has been ...provide faster access to a larger number of viral preps. Visit our Sponsors and Supporters Page Thank you...proud to be adding viral vector services to our repertoire. Additional Resources on the Addgene Blog ...

...use of both when making plasmids. Use repositories Repositories such as Addgene help share materials quickly...feels like an eternity of cloning, waiting, and repeating before you can finally dive deep into generating...everything correctly the first time – no mistakes, no repeats! How long does it take in total? While much harder...resources, and, on the rare occasion things go wrong, replace them for free. Plasmids ship out 2-3 days after...in a 96-well (or 384-well) plate! How much do repositories help?   Designing precise calculations for any...clear that community engagement with and use of repositories like Addgene provides large-scale savings. How...find plasmids Search for a plasmid in Addgene’s repository ...

...as T-DDOG and Cre-DOG, respectively, repurpose popular GFP reporter lines for more sophisticated experimental...a UAS-driven luciferase reporter construct. In the absence of GFP, no reporter output was observed in ... and a reporter construct into the mouse retina. GFP expression robustly activated the reporter gene TdTomato...GFP. They then tested T-DDOGs with two mouse GFP reporter lines; again, TdTomato was seen only in cells ...tested in retinal electroporation studies, DsRed reporter expression was induced in ~76% of GFP+ cells, ...

...particles and the virus particles produced could not replicate. However, Mother Nature found a way to re-create...packaging plasmid forming a vector that could create replication competent virus. Even though this system was ...still a potential risk for the generation of replication competent lentiviruses (RCL) especially if you...factors in HIV positive cells could make the virus replication competent. The 3rd generation of lentiviral...transgenes (>8 kb) Potential for generation of replication-competent lentivirus (RCL) Efficient gene transfer...transfer Potential for insertional mutagenesis: Replication-incompetent lentiviruses with human tropism can.... The addition of Dox in the culture will then repress your gene of interest. This system is efficient...

...used to create knockout mice include a reporter gene, which replaces the targeted gene and provides a way...genotype and phenotype and helps improve the reproducibility of experiments. Inbred mice are particularly... but because CRISPR/Cas9 engineering favors DNA repair via non-homologous end joining, the newly inserted...introduced to the nucleotide sequence during the DNA repair process. The addition of small molecules that promote...regions of the genome, and induce non-homologous repair at the site of DNA damage. These nucleases are ...embryonic stem cells. Also, because non-homologous repair is a random process, zygotes edited using the same...mediated activation of Cre to either induce or repress Cre expression using TetON or TetOFF systems. So...

...host genome, the virus is replication incompetent, meaning it can’t replicate and continue the infection... tools, thought was given on how to keep the replication of virus and its infectivity in check. Engineering...envelope plasmid), and the polymerase (Pol) for replication and Gag structural protein on yet a third plasmid... this safety feature prevents the virus from replicating and spreading in non-target cells, including ....    Viral cargo-specific safety   After replication competency, the second concern with virus safety...If administering virus to mice, part of your preparation should be doing your best to be well-rested and...familiarize yourself with your organization’s reporting mechanisms before you start an experiment, just...

... The FXP id also able to do high throughput DNA preps and other molecular work if we need it to. This ...Automating DNA purification We're developing ways to prep large quantities of DNA because some countries just...bacterial samples at the same time as opposed to preparing each sample individually. Enter the BIOROBOT! ...BIOROBOT Universal Platform and it suits our plasmid prepping needs perfectly. Working with Qiagen, we developed... the similar buffers as would be used for a mini prep, except it does it on a larger scale. Using the ...between samples. With this automated protocol, we can prep two 96 well plates in just under 3 hours or one ...You might not be doing enough transformations, minipreps, or filling enough vials to automate these processes...

...State Ladder: Homemade Plasmid Prep Penn State Ladder: Commercial Plasmid Prep Kit Amount 1000 Lanes 1000...significantly cheaper. When using a homemade plasmid prep protocol, it cost less than $10 to make 1000 lanes...bp or the 1 kb ladder. Using a commercial plasmid prep kit increases the costs to $20-30 per ladder: still...DNA ladder? You can find the Tan Lab’s DNA ladder prep protocol here and the pPSU1 and the pPSU2 plasmids...Generating DNA Molecular Weight Markers. Scientific Reports,7(1). doi:10.1038/s41598-017-02693-1. PubMed PMID...Addgene.org Use the Kit-Free Alkaline Lysis Plasmid Miniprep Protocol Learn How to Analyze Your Restriction...

...years at Addgene, we are sharing stories of our repository’s positive impact on science and scientists around...They did this by depositing them in Addgene’s repository. Other researchers, eager to try this new technology...their own to share. Over the years, Addgene’s repository has grown to include many CRISPR tools.  Distributing...Addgene had almost 18,000 CRISPR plasmids in our repository, and we have shared almost 300,000 CRISPR plasmids...of libraries, once made available through the repository. And the number of plasmids and libraries deposited...categorized by year of deposit. Outlines show a representative proportion of CRISPR plasmids requested, categorized...posts are also frequently read: Homology Directed Repair has been viewed 219,289 times, while Validating...

...unique skills and strengths (you will likely have reports with different levels of experience), find out ...tasks before I was a manager! Second, my direct report was not engaged because I only left him/her with...you’re delegating a recurring task to a seasoned report, there should be questions! Remember that delegating...itself is counter-productive. I always assume that reports want to succeed at work and providing feedback ...feedback is needed for that. In many cases reports are not aware of issues until you bring them up. I wouldn’...wouldn’t go as far as saying that your reports will be grateful for the feedback, but at least you can start...someone’s hard work and reward a job well done. Your report will (needs to) feel valued. Talk to your manager...

...resulting double strand break (DSB) stimulates DNA repair mechanisms that can be exploited to either inactivate...making targeted modifications to DNA, Cas9 can be reprogrammed to serve as a regulator of gene expression. ...domains such as transcriptional activators or repressors. These fusion proteins retain the ability to ...conventionally used SpCas9 (derived from the bacterium Streptococcus pyogenes). Consequently, fewer genetic tools...observed at the target site.   Figure 3: Gene repression with nuclease-competent Cas9. When complexed ...gRNAs targeting different sites of interest thus represents a straightforward means of exerting tight control...simultaneous cutting and transcriptional activation or repression at the respective sites.   Many thanks to our...

...species. Reproductive confinement involves working with laboratory organisms that can't reproduce with wild...wild-type chromosome and its own gene is used as a repair template, thereby converting the heterozygote into...experiments are less hazardous in organisms that seldom reproduce sexually because the drive must be much more efficient...confinement methods. Since molecular, ecological, and reproductive confinement typically require very little effort...an accidental gene drive release, which would represent proof positive that at least some scientists could...confinement are those of the author and do not represent a formal stance taken by Addgene or its staff....Insect Populations: I. Cage Studies of Chromosome Replacement by Compound Autosomes in Drosophila melanogaster...

...group dependencies  Take, for instance, a recent preprint released by the Boehm and Beroukhim labs at the...research uses ancestry groups rather than self-reported race, it highlights the myriad issues researchers...the results. But the reference genome does not represent the diversity of genetic variation across the ...theory, if the DepMap analysis had an equal representation of every ancestry group, they should have found...designed a tool that could analyze CRISPR sgRNAs and report the likelihood of any given sgRNA having an ancestral... researchers to ignore or miss. The lack of representation in commonly used cell lines led to an incorrect...complicating the issue is the long and ugly history of reporting false essentialist links between biology and ancestry...

...containing a phage origin of replication that can be packaged into replication-incompetent phage particles...antibiotic resistance is upon us, and the world is unprepared. Each year in the United States, two million ...essential processes: cell wall synthesis, DNA replication, or protein synthesis. Because they target such...sequences (red). Treating a mixed population, represented here by cells of different colors, specifically...small pools of antibiotic resistant cells could repopulate the environment after treatment. The phagemid...mammalian viral vectors in that the vectors are replication-deficient. Thus, a very large number of phagemids...CRISPR antimicrobials combines them with both replication-competent phages and antibiotics. Phage therapy...

...transcriptional repressors, they found that fusing dCas9 to an Mxi1 repressor domain repressed transcription... Covalent tethering of DNA repair template to Cas9 Homology-directed repair (HDR) can be used to make ...Gordon's lab developed a strategy to enhance HDR repair using a single-stranded oligodeoxynucleotide HDR...interference (CRIPSRi) platform for targeted gene repression in Candida albicans was developed by Rebecca ...

...Career Development; a Stakeholder Workshop Report.” The report enabled the development of priorities for...Development Hub Workshop Attendees and their 2021 report. Image from pd|hub.   To begin to tackle these...Steering Committee and working groups with representatives from all areas of the scientific training and...the findings were captured in this just released report “Enhancing Dissemination of Evidence-Based Models...programs.  Please join us for the pd|hub Workshop Report Public Release Event scheduled for March 4th from...2:30-4:00 pm ET.  You can learn more about the report's recommendations and join with us to discuss and...

...the new plasmids, antibodies, and viral preps in our repository through our Hot Plasmids articles.  Here's...facilitate SARS-CoV-2 research New viral vector preps of Voltron2 voltage sensors AAV vectors for targeting... tRNAs, within the lentiviral 3’ long terminal repeats (LTR). The 3’ LTR is duplicated during lentiviral...antibodies were recently added to the Addgene repository of tools for SAR-CoV-2 research. Two new antibodies...voltage sensors By Brian O’Neill New viral vector preps expressing the Voltron2 voltage indicator, created...-microglia cells.      Figure 5: A) Representative confocal images showing GFP expression from...Disease, including plasmids, antibodies, viral vector preps, CRISPR tools, and more.  ...

...experiments. If Cas7-11 lacked this toxicity, it would represent a major step forward for CRISPR-based RNA editing.... Specifically, efficacy rates of Cas7-11 were reported as up to 85%, broadly similar to shRNA and Cas13...However, in those same comparisons, the AbuGoot lab reported no detectable effect on cell viability – meaning...mutations wouldn’t matter – the mutated copy could be replaced by simply dragging and dropping in a functional...allowing a one-size-fits-all approach to gene replacement therapy for most patients. But how does one go...work. (It was also found to be independent of gene repair.) This lowers the chance of the insert being “dropped...techniques like HITI or HDR, which require specific repair pathways to work, PASTE is more specific and efficient...