We narrowed to 798 results for: REP

...regulatory elements (to mutate promoters/enhancers in reporter constructs), in functional analysis of proteins...introduced to endogeneous DNA through homology-directed repair (HDR) of a CRISPR/Cas9 induced double-stranded ...to the endogenous target, yet to facilitate the repair, the template should be resistant to Cas9 cleavage...your primers should be free of palindromic and repetitive sequences, but if present, a minor extension ...product (see figure). Template A high purity plasmid prep significantly increases the success rate of site...’ll want to steer clear of these strains when preparing the plasmid template for site directed mutagenesis...

...versions obsolete, the techniques we use in lab are replaced by improved versions that save both time and money...observed. Today Southern blotting has been largely replaced by real-time PCR to answer the same experimental...researcher (or a simple computer program) can then reproduce the DNA sequence (Figure 2). Accuracy and accessibility...this technology, radiolabelled nucleotides were replaced with fluorescently labeled ones allowing for fluorometric-based...discoveries (5, 8). While newer techniques have largely replaced these, these techniques still have useful applications...already commercially available although they are reported to have high error rates (10). It is projected...

... our viral vectors repository this year (and it's only March!)  In our repository, you'll find the latest...What's new in Addgene's ready-to-use viral vectors repository? Quite a bit! In this post, we'll share the 21...New ready-to-use viral vectors in the Addgene repository  Name Serotype Category Depositor  Notes ...check out Addgene's Data Hub to view viral vector reports from other users. And don't forgot to use the link...

...research" (2013) Scientific Reports 3, Article number: 1649 doi:10.1038/srep01649    ... to all articles, Anurag Acharya and colleagues report, the percentage of citations to articles in non-elite...publications. (As an interesting aside, Nature recently reported that the most cited work in history is a 1951 ... the best way to measure impact? As that Nature report also noted, papers detailing Nobel Prize-winning...

...gradually diminish over time; and, importantly, repeated excitation generating reactive oxygen species ...difficult-to-study genes. Its main function is to regulate DNA repair, senescence, and apoptosis. Work by Kamio et al...the authors used a p53 response-element NanoLuc reporter to show that the mutants abolished transcriptional...This work highlights HiBiT tagging as a sensitive reporter for tracking proteomic cargo flow via EVs. Find...., & Wood, K. V. (2012). Engineered luciferase reporter from a deep sea shrimp utilizing a novel imidazopyrazinone...carcinoma in human tissue and in vitro. Oncology Reports, 50(5). https://doi.org/10.3892/or.2023.8631 Tsien...

...plasmid" section Anytime you use a plasmid from the repository, we request that you cite it using both its Addgene...find the fruits of your labor and won’t have to repeat the modification. This will save both time and ...you to browse through the many plasmids in the repository, our scientists have curated a number of special...attention to the great variety of plasmids in the repository. And last, but certainly not least, we often ...Special Collections Pages Suggest Plasmids for the Repository Find Plasmids for Your Research  ...

...before using an antibody.  Antibodies and Reproducibility First, let’s review why validation is so important...called out antibodies as major culprits in the reproducibility crisis of biomedical research (Baker, 2015;...proposed 5 Pillars of antibody validation which represent five experimental strategies researchers can use...benefit and we can all help improve antibody reproducibility!     References and Resources Additional ...plasmids here!  References Baker M (2015) Reproducibility crisis: Blame it on the antibodies. Nature ...10.1038/521274a  Bradbury A, Plückthun A (2015) Reproducibility: Standardize antibodies used in research. Nature...

...behaving mice to gather the following types of data: Report time-course and concentration of released DA in...optogenetically activated and inhibited neurons in live mice Report modulation of DA signaling during learning and ...moving mice to gather the following types of data: Report endogenous DA release from a single dopaminergic...looming stimulus in vivo in transgenic zebrafish Report DA release in live mice over the course of Pavlovian...of our viral vectors blog posts Find tips for preparing AAVs Download our viral vectors 101 eBook Resources...

...of a plasmid, the first step is always to send prepped plasmid DNA for next generation sequencing. We ...promoters, selectable markers, tags, origins of replication, and fluorescent proteins that are already included...sequence by NGS. These can include GC-rich and repetitive regions. These regions require additional verification... plasmid off for Sanger sequencing.     If a repetitive region is present in the plasmid, we might need...analyze sequencing results for plasmids in the repository at a glance. Our Snapgene-powered plasmid maps...

...ADAR constructs from the REPAIR (RNA Editing for Programmable A to I Replacement) system. Like RNA targeting...Matchmaker Day 2: Homology directed repair: CRISPR 101: Homology Directed Repair and 3 Tips to Improve HDR Efficiency...Requirement and Expanding CRISPR Beyond SpCas9 Day 8: Repair mechanisms: CRISPR 101: Non-homologous End Joining...

... Specifically, viruses are able to infect and replicate in host cells, but they are susceptible to cells...transposon consists of a mirrored set of inverted repeats (green) flanking the gene of interest (blue) in...is expressed (red stars) and binds the inverted repeats (green); and an endonuclease reaction occurs which...puromycin, hygromycin) and fluorescent protein reporters (GFP, BFP, RFP, no FP). The vectors each contain...region of the RPL41 gene, to drive the selection/reporter parts of each construct. This new tool kit allows...

...plasmid. Inverted terminal repeat (ITR) sequence Inverted terminal repeats (ITRs) are what makes an AAV...contains the proteins needed for the virus to replicate,  and the transfer plasmid which contains the ...’s ~4.4 kb or smaller. Some examples include a reporter like GFP, some smaller CRISPR-Cas proteins such...(2019) A User’s Guide to the Inverted Terminal Repeats of Adeno-Associated Virus. Human Gene Therapy Methods...

... This allows them to replicate viral plasmids with the SV40 origin of replication. To generate a virus...considered hypotriploid: most chromosomes are represented by two to three copies, but several have four...your research question, HEK-293 is known for reproducible results. It’s also widely available, which can...biology, ion channels, or involve human protein preparations with PTMs. Why the HEK not? References and ...

.... In doing so, the dimer shifts from one that represses transcription to one that activates transcription.However...BLADE The team modified AraC to create BLADE by replacing its dimerization domain with VVD, a light-responsive...the DNA-binding domain of AraC using a mCherry reporter downstream of a PBAD promoter. While none of the...contains BLADE, the PBAD promoter, and a mCherry reporter downstream of the PBAD promoter. To use this plasmid... plasmid with your gene of interest, you would replace mCherry with your gene of interest. pBLADEONLY:...

... posts of 2015 1. CRISPR 101: Homology Directed Repair Addgenie Chari Cortez explains some of the basic...basic science behind homology directed repair (HDR) and how it’s used in CRISPR genome editing.   2. Plasmids...While the vast majority of CRISPR plasmids in our repository use Cas9 to cut DNA, Addgene Science Communications...explain some of the molecular gymnastics behind DNA repair by non-homologous end joining (NHEJ). 7. Career...Resources Learn More about Viral Vectors Browse the Repository  ...

...You’ve prepped your DNA and you’re ready to get started on the next step of your experiment. But in many...give you an idea of the concentration of your DNA prep and whether there are any other contaminants. Nucleic...your sample against this curve to quantify your DNA prep. Though this method takes more time to set up in...and is not used as much for the standard plasmid prep. Diphenylamine method Another absorbance based method...Learn about the different grades of plasmid DNA preps Resources on Addgene.org Visit the Molecular Biology...

... about sexual and reproductive health, as the stigma around sexual and reproductive health issues creates...to all the optional sexual health sessions and reported the intel back to my shyer classmates. For my ...evaluate the quality of the study those abstracts represented. The point of this study was to see if their ...research focuses on misinformation about sexual and reproductive health.    References and Resources References...

...Gerald Schwank, first author of the cystic fibrosis report. “This is a big advantage over induced pluripotent...also need to be perfected until the efficiency of repair approaches 100 percent. “We need to be sure it'...it's accurate enough and efficient enough for repair,” Doudna said. “And it can't be too hard to apply....need to get a handle on how cleavage sites are repaired.” Those insights will be critical for scientists...December 2013). 8. Gerald Schwank et al. "Functional Repair of CFTR by CRISPR/Cas9 in Intestinal Stem Cell ...

...enough to get it into cells and let them do the repair work. The key is T4 DNA polymerase, which has 3...forming a circular product with four nicks easily repairable by the bacteria after transformation. LIC is ...transformed into bacteria, which will efficiently repair the plasmid. Created with BioRender.com.   ...intermediate is transformed into bacteria, which repair the plasmid. Created with BioRender.com. How...PCR-generated inserts. By harnessing the power of DNA repair, you can assemble multiple fragments without the...

...convinced that most microscopy techniques can be replicated with commercially available consumer-grade components... tools has allowed the community to regularly replicate designs worldwide. Distributing hardware via globally...Detailed documentation, which facilitates the reproduction of the structure in a similar way to the biological...assemblies open-source. The sharing of digital representations of the 3D models, which in many cases can be...with the goal of creating more transparent and reproducible science in the long run (Hohlbein, et. al., ...