We narrowed to 791 results for: REP

...the new plasmids, antibodies, and viral preps in the repository through our Hot Plasmids articles.  Here's...delivered with an enhanced pU6cm promoter, viral replicon amplification of cargo, and heat treatment.  ...//doi.org/10.1038/s41477-024-01786-w. (bioRxiv preprint: https://doi.org/10.1101/2024.02.11.579803).  ...components of many physiological pathways and represent the largest family of human drug targets. To enable...6E10-A5] in Immunohistochemistry (Human). Addgene Report, doi: https://doi.org/10.57733/addgene.6ncijt.  ...

...multiple other labs have reproduced their EGFP findings since the initial report in 2023.)     Figure.... Andrew and Maria were encouraged by previous reports that described magnetic responses of cryptochrome...indicate times when the magnetic field is on. Image reproduced from Frank Hayward et al. 2024 under CC-BY license...), its behavior has already been successfully reproduced by several other labs. Andrew and Maria expect...

...technology that allows precise modifications of DNA (replacements, small insertions, and deletions) without introducing... In our study, we used the TRIP (Thousands of Reporters Integrated in Parallel; Akhtar et al., 2013; Schep...Chromatin Position Effects Assayed by Thousands of Reporters Integrated in Parallel. Cell, 154(4), 914–927...., Raguram, A., & Liu, D. R. (2019). Search-and-replace genome editing without double-strand breaks or ...insertion efficiencies using sequence features and DNA repair determinants. Nature Biotechnology 2023, 1–11. ...context on Cas9-induced DNA double-strand break repair pathway balance. Molecular Cell, 81(10), 2216-2230...

...the sequencing results of the plasmids in our repository with known reference sequences, such as full ... entries in NCBI’s numerous databases. As our repository has grown over the years (we now have over 60,000.... At Addgene, we use blastn to identify any discrepancies in Sanger sequences, such as mismatches, deletions...determine the potential effect of any nucleotide discrepancies. The blastp and blastx programs are optimized...Similar to nucleotide sequences, proteins often have repeated or highly homologous regions, which by default... standard blastx search. An alignment omitting repeated regions can be confusing, such as when you attempt...

..., consider an approach that prioritizes more reproducible/renewable reagents, such as recombinant or hybridoma-derived...cells that are amenable to genetic knockout, this represents a very robust approach to antibody validation...quantified using the Attune NxT Flow Cytometer with representative images showing the staining intensity in the...increase confidence that the labeling observed represents the expression level of the target of interest...antibody clones The antibody validation process represents a high level of effort and time, which may not...evidence is then needed to confirm that the staining represents the target in your sample of interest (bottom... specific detection of human TRPA1. Scientific Reports, 9(1), 18500. https://doi.org/10.1038/s41598-019...

...purification kits. Kit-free alkaline lysis plasmid miniprep Starting material: 2 mL bacterial culture Product...Product: plasmid DNA This kit-free plasmid miniprep protocol from Addgene follows a similar workflow as ...authors estimate that a gravity-based column plasmid maxiprep kit takes 1-1.5 hours to complete, while this ... minutes. Additionally, while commercial plasmid prep kits come in discreet sizes (mini, midi, maxi, giga....0007750  Vogelstein, B., & Gillespie, D.A. (1979). Preparative and analytical purification of DNA from agarose...Purification and Gel Extraction Kits. Scientific Reports. https://doi.org/10.1038/s41598-018-30316-w  Zou...

...transformation   Spectinomycin/streptomycin Spectinomycin and streptomycin both inhibit protein synthesis...expensive antibiotic compared to streptomycin. Spectinomycin/streptomycin can be successfully used for plant...offers is marginal. Fortunately, amp can easily be replaced with carb if satellite colonies are an issue. ...spectinomycin). Cons: Cost (spectinomycin) or stability (streptomycin), doesn’t work for all bacteria.   Zeocin  Zeocin...

...transcription) is demarcated by viral genome replication (Harrach, 2014). Given that the early genes ...encapsidate new virions) are not expressed. Note that replication-competent AdV vectors also exist and are used...the final topic, which is that natural AdVs, replication-competent viruses with a recombinant genome, ...(Tollefson et al., 1996) – cell death has been reported when using vectors, and immune inactivation of...pre-existing toolbox for adenoviral vectors in Addgene’s repository is smaller than for other vector types, we hope...Hermiston, T. W. (2003). Multigene expression from a replicating adenovirus using native viral promoters. Mol ...

...activator such as VP64 (Gilbert et al., 2013), or repression of transcription through binding a promoter region...expression plasmid. Experimental use Since the initial report describing CRISPR-mediated purification of a specific...dCas9, with bound DNA, can then be purified by streptavidin high-affinity purification. This system is both...including Protein A, 1xFLAG, 2xAM, and HA have been reported, as well as affinity purification with an anti-Cas9...chromatin immunoprecipitation (enChIP)." Scientific reports 3 (2013): 3171. PubMed PMID: 24201379. PubMed Central.... PubMed PMID: 28841410. Qi, Lei S., et al. "Repurposing CRISPR as an RNA-guided platform for sequence-specific...

...nuclease-deactivated Cas9 (dCas9) can be tethered to a repressor (CRISPRi) or an activation domain (CRISPRa) to...CRISPRko sgRNA library as pooled library or lentiviral prep! Dolcetto: human CRISPRi sgRNA library While CRISPRko... protein (dCas9). dCas9 is then tethered to a repressive domain, such as KRAB, to prevent efficient transcription...Dolcetto library! Now also available as a lentiviral prep. Calabrese: human CRISPRa sgRNA library In addition...Find the Calabrese pooled library and lentiviral prep here! Applications for the Root and Doench CRISPR..."Genome-scale CRISPR-mediated control of gene repression and activation." Cell 159.3 (2014): 647-661. ...

... in cancer studies, act as a reporter for DNA double strand break repair, and to label pathogenic intracellular...maintaining fluorescence. The protein structure, first reported in 1996, is an eleven β-sheet-containing “barrel...been done to expand the red-fluorescent protein repertoire; however, these proteins are unique from GFP ...constructing fluorescent fusions. Transcription Reporter: Placing GFP under the control of a promoter of...a given cell type. This type of transcription reporting was among the earliest uses of GFP. Fluorescence...

...the new plasmids, antibodies, and viral preps in the repository through our Hot Plasmids articles. Today... set of fusogenic and targeting components to reprogram delivery vehicles like lentiviral vectors, CreVLPs...doi: 10.1038/s41592-023-01880-5. PMID: 37188954. Preprint available on bioRxiv – doi: 10.1101/2023.03.09.531939...doi: 10.1016/j.cell.2023.05.044. PMID: 37385249. Preprint available on bioRxiv – doi: 10.1101/2023.02.07.527548...doi: 10.1038/s41587-022-01524-7. PMID: 36593408. Preprint on bioRxiv — doi: 10.1101/2021.10.13.463862.  ...

...in our repository that you can use to study these genes! Groucho, a transcriptional co-repressor The first...after the protein Groucho, a transcriptional co-repressor important in development. Groucho family proteins...specific set of DNA-binding transcription factors to repress the expression of genes primarily involved in development... see for yourself. We have one plasmid in our repository, plasmid 39624, that contains cmr.   tinman and... Groucho/TLE/Grg family of transcriptional co-repressors." Genome biology 9.1 (2008): 205. PubMed PMID...

... strand break (DSB) mediated homology dependent repair by enhanced green fluorescent protein (EGFP) reconstitution...sgRNA guided Cas9 endonuclease, homology dependent repair (HR, homologous recombination; SSA, single strand...halves of EGFP to recombine by homology directed repair, and resulting in the expression of EGFP. Using...Samantha Young is a Medical Writer with a PhD in reproductive biology and genetics. She loves to communicate...expressing Cas9 and single guided RNA." Scientific reports 3 (2013). PubMed PMID: 24284873. PubMed Central...Non Homologous End Joining and Homology Directed Repair Additional Resources on Addgene.org Find Software...

...Transfer Plasmid : 1 pHelper: 1 RepCap OR 3 Transfer Plasmid : 5 pHelper: 2 RepCap Select what type of tissue...goes, “failing to plan is planning to fail.” Preparation is a key step to any experiment and can help ...experimental details on the spreadsheet: Name your viral prep Enter the specific names of the plasmids you plan...using 28 μg of total plasmid (Transfer, helper, and RepCap added together) per 15 cm dish or 2500 μg of plasmids...

...dCas9-VP64) or repressors (dCas9-KRAB or dCas9-SRDX) can also be used to activate or repress target genes...engineering CRISPR can be used to knockout, activate or repress target genes in plants using the same general experimental...containing plasmids for knockout, activation and repression of target genes in plants and many of the aforementioned...in human cells. In contrast to homology directed repair, these methods do not involve double strand break... the Patron Lab.  As with all plasmids in the repository, we highly recommend reading the associated publications...

...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...

...similar protein that was discovered in the 1960s, strepavidin. The Biotin-avidin complex can then be used and...primary antibody respectively. Avidin bound to a reporter is then added to the reaction for easy detection... experiments. During detection, a biotinylated reporter enzyme that is pre-incubated with free avidin ...protein of interest leading to amplification of the reporter signal and easier detection. This strategy is ...Forster, A. C., Skingle, D. C., & Symons, R. H. “Preparation and uses of photobiotin.” Methods in Enzymology...mammalian cells with biotin ligase and monovalent streptavidin.” Nature Protocols 3.3 (2008): 534–545. PubMed...

... 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...