We narrowed to 791 results for: REP

...vector preparations, we have used quantitative PCR to evaluate the physical titer of our preps. We often...often repeat this assay to confirm that our results are accurate. But what if we could repeat this assay... for ddPCR set-up. Prepare the master mix in a separate area from where you prepare your sample dilutions...detection of the product. As an added benefit, replicates are built into the technology. One well (containing...

..., Zetsche et al. hypothesized that Cpf1 might represent a functional CRISPR nuclease that could be adapted...about the same size as Cas9’s, but with the direct repeat preceding the spacer rather than following it. ... only a short stem-loop structure in the direct repeat region is necessary for cleavage of a target. Cpf1...are difficult to modify through homology-directed repair (HDR). Cpf1 also expands the number of sites that...experimentally confirmed. To prevent new editing post-HDR, repair templates should remove the PAM sequence. Comparing...in mammalian cells, LbCpf1 and AsCpf1. In both reports, on target editing efficiency for the Cpf1 orthologs...mismatches, but variably so, with Kleinstiver et al. reporting that Cpf1 can tolerate mismatches at gRNA positions...

...these proteins, as well as the viral long terminal repeats (LTRs) are different. As a result, lentiviral and...adenoviral protein E1A, which has been shown to repress expression of HIV-1 LTRs (more information is available... thus decreasing the probability of creating replication-competent lentivirus (RCL) in your target cells...needs to be targeted. Q6: Are transfer plasmids replication competent? What is SIN? A6: Most (if not all)...all) Addgene transfer plasmids are replication-deficient meaning they can be used to create virus that is...Which genes are deleted or modified to result in replication deficiency? What percent of the vector components... vectors are: 1) the potential generation of replication competent virus [usually HIV-1]; and 2) the potential...

...different CGBE tools allow the researcher to direct the repair of the abasic site. But what if an editor could...excision is the most common type of base excision repair. But mammals also have an enzyme to remove inosine...collaborative team from the David Liu and Dali Li labs reported a similar tool shortly thereafter, using mouse... or to create the substrate for a base excision repair pathway. The team behind AYBEv3 introduced the ... the non-edited strand to encourage the cell to repair the DNA using the edited strand as a template —...editors Excises G, T, or C, respectively, to be replaced with another base. References Chen, L., Hong...Transversion Editors Read about four base editor reporters Resources on Addgene.org Browse CRISPR plasmids...

...jejuni (Cj) NNNNRYAC Streptococcus thermophilus (St) NNAGAAW Treponema denticola (Td) NAAAAC...in mammalian cells. The REPAIR (RNA Editing for Programmable A to I Replacement) system fuses Cas13b to...when trying to edit a gene using homology directed repair, since HDR-mediated gene editing is most efficient...Hu et al., 2018).  In 2020, the Kleinstiver lab reported the development of near-PAMless Cas9 variants....below.  Cas9 species PAM sequence ’) Streptococcus pyogenes (Sp) 3' NGG Staphylococcus aureus...

... organisms) -  perhaps you’ll choose to study Crepidula atrasolea or Prochlorococcus with your next project...used to study aspects of cognition. Spiralians - Crepidula atrasolea (Jonathan Henry) This black footed slipper...real bottleneck. New gif for #MolluscMonday: #Crepidula atrasolea juvenile, light microscope with polarizing...it in a fertilized egg to get edited embryos (replacing the nucleus is still very difficult). Cuscuta ...targeted gene mutations in anoles lizards or in any reptiles at all. But new success is coming with CRISPR/...full hydra and hydra have asexual and sexual reproductive life cycles. One can inject plasmid DNA in the...adult female mosquitoes. Now they are trying to replace embryo injection (hard to do, expensive, not every...

...express my concerns and request a replacement. I got my replacement in the end and the issue was resolved... A degree in the life sciences prepares one for numerous non-academic careers. Still, many start their...support roles. Examples are Technical Service Representative, Technical Support Specialist, Technical Support...less suitable for troubleshooting or issuing replacement products. In order to find good and helpful answers...certainly there won’t be a shortage of amusing reports where someone tries to “optimize” a particular ...job. Solutions may also be achieved by simply replacing the product in question. This can solve the problem...

... to conduct experiments faster, skipping the preparation steps of production, purification, and titering...customizability, choosing the cargo, serotype, and prep size they need for their experiments — and we’ve...them out in record time.  However, our in-stock preps remain a popular choice thanks to the convenience...cortical interneurons. Take a look to see if these new preps might enhance your research!   Table 1: New ready-to-use...ready-to-use viral vectors in the Addgene repository  Name Serotype Category Depositor  Notes pAAV-CAG-emiRFP670...help, with over 100 user-submitted viral vector reports. And don't forget to use the link below to keep...

...What's new in Addgene's ready-to-use viral vectors repository? Quite a bit! In this post, we'll share the 20... 20 additions we've made to our viral vectors repository since March.  Addgene is now offering an AAV2...gene expression! We've also added AAV1 and AAV5 preparations of several variants of a new optogenetic tool...Yizhar lab. Addgene is now offering AAV1 and AAV5 preparations of several variants of a new optogenetic tool...New ready-to-use viral vectors in the Addgene repository  Name Serotype Category Depositor  Notes ...check out Addgene's Data Hub to see viral vector reports from other users. And don't forget to use the link...

...gradients? Excellent! Now, let’s head west, intrepid reader! Protein Prep The first step in any western blot is...minutes) at 37 °C, if you suspect aggregation in your prep. If your proteins are the wrong size on your gel...will simply load an extra ladder or technical replicate (ie, leftover sample) to fill the relevant wells...antibody’s dynamic range. A) The protein dilutions, representing expected concentrations, are all within the ...antibody’s dynamic range. B) The protein dilutions, representing expected concentrations, are outside of the ...blot, you may want to consider using a different reporter or, if you don’t need to compare the protein amounts... amounts directly, stripping and reprobing. Just make sure to go from least to most abundant protein if...

...fluorophore-conjugated streptavidin protein, which binds the primary antibody through the biotin-streptavidin interaction...detection. The advantage of this system is that streptavidin conjugates are commercially available for a ...fluorophore selection for panel design. Find streptavidin plasmids and biotin plasmids at Addgene!   ... two-protein labeling by utilizing(a) biotin-streptavidin binding and (b) antibody host species interactions...include transcription factors, histones, and DNA repair enzymes. Note that when using the nuclear fix/perm...or protein-protein interactions, like biotin-streptavidin binding and antibody host species reactivity...

...talk to the occasion Don’t just plan to reuse a prepared talk unless it perfectly fits the occasion.  I...figure out how to make it suspenseful. 4. Start preparing your talk early - like today Don’t wait until ...for poor quality slides with sufficient time to prepare (and it does take time). For important talks, ask...loves a great animation. 8. Predict questions, prepare answers Speaking pros always have a few extra slides...come up with clear and concise answers. This preparation will help you avoid giving only yes or no answers...their phones. Speaking longer just makes you seem unprepared or arrogant. A bit short is always better than...when I then ask “and who has deposited to the repository?” When most or all of the hands go down it is...

... are overrepresented on Twitter Mathematical, life, and physical scientists are underrepresented. It’s...interested in getting started? Reply in the comments below or tweet us your reply & #scientistswhotweet. And...users. Biologists rank #8. Figure 1: Highly Represented Scientific Disciplines on Twitter. Twitter users...Follower Network of Scientists on Twitter. Each dot represents a scientist. Only scientists who follow each ...

...serotypes at Addgene. Serotype In-stock viral prep Packaged on Request AAV1  ✓ ✓ AAV2 ✓ ✓ AAV2... targeting astroglia. It has additionally been reported to have tropism for liver, vascular endothelial... muscle inflammation and degeneration has been reported (Muraine et al., 2020). AAV8 AAV8 has high transduction... the CNS (Han et al., 2023), and has also been reported to have tropism for liver and kidney cells (Issa...Use Addgene’s AAV Data Hub to find experimental reports on commonly used vector cargo designs and administration...cells utilizing various AAV serotypes. Scientific Reports, 15(1), 20927. https://doi.org/10.1038/s41598-025...AAV Packaged On Request Browse all in-stock AAV preps Browse all AAV plasmids ...

...engineered not to cleave DNA and instead activates/represses gene expression by various mechanisms. The RNA...cleaving of DNA occurs, not for deadCas activator/repressor experiments where only localization of the Cas...will generate a ssDNA break which can easily be repaired by cells with a near zero likelihood of mutation...insertion of DNA by the NHEJ pathway during DNA repair of the break to allow for site-specific detection... these approaches is that NHEJ must perform the repair to allow for detection, not Homologous Recombination...non-cutters If your Cas experiment involves activation/repression or any other effect which does not depend on ... be an appropriate method for an activator or repressor Cas. Similarly, Cas ChIP-Seq would also identify...

...the new plasmids, antibodies, and viral preps in the repository through our hot plasmids articles. Think...pitch a Hot Plasmids section (antibodies and viral preps welcome!) here. Here's what you'll find in this...Liu Z, et al. Cell. 2022.  Find JEDI-2p viral preps here (and here!)Find JEDI-2p plasmids here!  Liu...adapted from Govorunova et al. 2022.     HcKCRs represent a long-sought tool for researchers to control ...s41593-022-01094-6. (This work was also shared as a preprint: https://doi.org/10.1101/2021.09.17.460684.)   ...

...data repository for validation data on Addgene/CLOVER viral vectors.  Caspedia: A data repository for ... Calculator app for iPhone Reagent and Data Repositories These links can help you find experimental and...for Cas enzymes.  Open repository of CRISPR screens Cellosaurus: A knowledge resource for cell lines IBEX...IBEX Knowledge Base: A data repository for validation data on IBEX reagents.  mAb3D atlas validated antibodies...human, and marmoset brain IHC.  FPbase: A data repository for fluorescent proteins and their properties...

... ReFigure. Reading and exploration including replications and experiments resulting in “negative data”...performance as in this antibody ReFigure Analyzing replications as in this ReFigure about whether Natronobacterium...reagent performs well, or you might publish a replication experiment that you conducted as part of setting...traditional publications. By submitting your data to a repository (Figshare is currently supported by ReFigure ... specialized knowledge of which findings are reproducible. For an example, see our ReFigures on the therapeutic... interim research products (pre-registration, preprints, data, software, micropublications) in grant proposals...

...in the specific repositories (Addgene, WormBase, Arabidopsis.info). These repositories will ensure that...published work meets the accepted standards of reporting in the discipline and reduces the dissemination...information that would prevent the study to be replicated or omission from the literature. I think that...data on ... The language is ... The general representation of data is... The figures are ... In general...the data can be deposited in the public on-line repository (like GitHub, Zenodo, Figshare, Triad) while ...

...efficiency. If this finding is replicated in larger studies, it will represent a key advantage over Cas9, ... RETRACTED AND FOLLOW UP STUDIES HAVE FAILED TO REPEAT THE RESULTS DISCUSSED BELOW Biologists are going...the question on everyone’s minds is: could NgAgo replace CRISPR? Such a drastic shift won’t happen overnight...NgAgo is also able to mediate homology-directed repair when a template is supplied. Find Plasmids from..., this property could be an advantage. However, repair after base excision would alter the target sequence...