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We narrowed to 1,038 results for: LAS

Showing: 201 - 220 of 1038 results

  1. We're Updating Our Hold for Publication Policy

    Type
    Blog Post
    ...will release plasmids on hold if they are clearly described in a journal publication. Plasmids will be released... and your deposit or plasmid ID numbers and we’ll be happy to update the plasmids to add the citation ...and unpredictable. Fortunately, you can submit plasmids at any point in that process and we provide an...necessary quality control and legal approvals so the plasmids are ready for distribution as soon as the paper...better addition to peer review than having your plasmids sequenced and openly registered for other scientists...put in us when you choose this status for your plasmids. In fact, almost half of all new deposits start...hold! You will always have control over when your plasmids are distributed. Next month, we are making two...

  2. Tips for CRISPR Gene Editing in Mice

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    Blog Post
    ... target plasmid and pX330-sgRNA plasmid (Mashiko et al. 2013). The pCAG-EGxxFP target plasmid contains...pCAG-EGXXFP plasmid along with the individual gRNAs in their PX330 plasmids (or similar plasmids for gRNA... (c) The pCAG-EGxxFP target plasmid was co-transfected with pX330 plasmids containing sgRNA sequences ... system floating around, and many pre-designed plasmids containing these variants ready for purchase. ...multi-cloning site (MCS) between EGFP fragments. The pX330 plasmid contains humanized Cas9 expression cassette and...HEK293T cells. The fluorescence intensity was classified into four groups (4; brighter than control, 3... you can insert this region into a pCAG-EGXXFP plasmid using standard cloning techniques (The XX indicates...

  3. Overcoming the Challenges of Lentiviral Production

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    Blog Post
    ... multiple plasmids. Second generation packaging systems use 3 plasmids; an envelope plasmid that usually...encodes VSV-G, a packaging plasmid containing structural genes and a transfer plasmid carrying the transfer...Rev gene from the packaging plasmid and incorporating it into a fourth plasmid. In addition, the promoter...and packaging plasmids in order to obtain the best titers. Requiring an additional plasmid for transfection... 5-layer flask will require some initial optimization; cell number, media volume, and plasmid concentrations...By dividing the viral elements across multiple plasmids, the risk of recombination and consequently, the...extra level of safety, they also require a fourth plasmid. The requirement for additional DNA can impact ...

  4. Popular Retroviral Vectors and Their Uses in Scientific Research

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    Blog Post
    ...separate envelope and packaging plasmids as well as transfer plasmids. This concept ensures safe handling...gamma-retroviral packaging plasmids are not interchangeable. General envelope plasmids, such as VSV-G, may be...Learn more about CRISPR plasmids for transcriptional inhibition. Retroviral Plasmid Kit Retroviral Barcoding...expressing new shRNA sequences pLKO (Plasmid #10878) This plasmid has been used by The RNAi Consortium... genes and transgene expression across several plasmids. The most commonly used viral vector systems are...popular packaging and gamma-retroviral envelope plasmids here. Lentiviral vectors are very versatile tools...guide. Popular packaging and lentiviral envelope plasmids are listed here. Please note that both lentiviruses...

  5. London Calling - Addgene’s New Europe Office in London

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    Blog Post
    ...Addgene, plasmids, and depositing (all while our US counterparts are still sleeping). Plasmid sharing ...more than 75,000 plasmids to more than 1,500 different institutions in Europe. In the last four years alone...have deposited their plasmids into the Addgene repository. More than 4,500 plasmids have already been deposited...in research, I developed several new plasmids, including: plasmids to study the dimerization of HLA B27... Years of Plasmid Sharing: Interested in more statistics from Addgene's 10 years of plasmid sharing? ...Check out our Top 10 Plasmid Technologies blog post Or look at our "10 Years of Plasmid Sharing" Infographic...emailed. And Addgene has been there to answer your plasmid questions. But until now, you’ve always had to ...

  6. CRISPR 101: Epigenetics and Editing the Epigenome

    Type
    Blog Post
    ...modifications, researchers used blunt tools like histone deacetylases, but targeted epigenetic modification was impossible... constructs, which fused a TALEN to the Tet1 demethylase catalytic domain, could mediate demethylation...additionally fused a TAL effector to LSD1 histone demethylase to demethylate enhancer regions (Mendenhall et...light-regulated histone methyltransferases and deacetylases. CRISPR and epigenetics As with many TALEN-based...activation. Created with BioRender.com. Tet1 demethylase Ronggui Hu’s lab has created pdCas9-Tet1-CD for...cytosine demethylation in mammalian cells. This plasmid is used with pcDNA3.1-MS2-Tet1-CD to decrease methylation...Tet1CD, is available from Rudolf Jaenisch’s lab in plasmid form or as ready-to-use lentivirus. Figure...

  7. CRISPR 101: Engineering the Plant Genome Using CRISPR/Cas9

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    Blog Post
    ... consists of a Ti plasmid, into which the GOI is inserted, and a Ti "helper" plasmid, which contains Vir...for creating plant expression plasmids, including plant CRISPR plasmids from the Yiping Qi lab and the...Originally published Oct 11, 2016 and last updated Dec 22, 2020 by Benoit Giquel. CRISPR has taken the...plant-specific modifications to commonly used CRISPR plasmids are necessary to use the CRISPR system in plant...et al., 2013). Addgene carries Cas9-containing plasmids for knockout, activation and repression of target...expression of both Cas9 and the gRNA off of the same plasmid. Delivering CRISPR components to plant cells Once...the application of this approach is limited to protoplast cells (plant cells whose cell wall has been removed...

  8. Sharing Your Lab Protocols: Using Apps to Save Time & Track Your Experiments

    Type
    Blog Post
    ... requested plasmids. I wondered how many scientists have used the protocol for this plasmid, and if it... your protocols onto your plasmid page, scientists who request your plasmid will have easy access to your...restriction digest of plasmid DNA, and bacterial transformation. And more plasmid-related educational resources...be found in our Plasmids 101 blog series. Want to learn more about sharing your plasmids via Addgene? Check... resources, such as protocols, Plasmids 101 articles on various plasmid features, and reference tables...deposit their plasmids to the repository. These protocols show up on the related plasmid pages and can... protocol is being used around the world? In the last month it’s been viewed by users in the US, China...

  9. Lentiviral Vector Uses and Overview

    Type
    Blog Post
    ...consisted of three plasmids: the packaging, envelope, and transfer plasmids. The packaging plasmid carried a mutated... packaging and envelope plasmids are two of Addgene’s most requested plasmids! Lentiviral particles with...Many CRISPR plasmids are designed for lentiviral use, including Addgene’s top requested plasmid in 2015: ...itself due to a few missing proteins. The envelope plasmid contained a viral envelope to dictate the tropism...the vector could infect. Finally, the transfer plasmid encoded the desired transgene flanked by HIV-1 ...genome integration. After co-transfection of these plasmids, 293T cells released transgene-containing lentiviral...recombination between the transfer and packaging plasmids does exist. To further reduce this possibility...

  10. Viral Vectors 101: Gamma-Retroviral Packaging Systems

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    Blog Post
    ...γ-retroviral packaging plasmids are not interchangeable. General envelope plasmids, such as VSV-G (Vesicular...in trans by specific plasmids. This method, therefore, reduces the number of plasmids that are required ...encoded by the transfer plasmid, but instead are provided in trans by other plasmids or the packaging cell...do many γ-retroviral plasmids have similar names? The nomenclature of the plasmid backbone is meant to...components are needed: γ-Retroviral transfer plasmid encoding a transgene, sgRNA, or shRNA of interest..., which facilitate integration of the transfer plasmid sequences into the host genome. Typically, it is... genome upon viral transduction. Most transfer plasmids available from Addgene were derived from MoMLV...

  11. REPLACR Mutagenesis: Replacing In Vitro Recombination Methods

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    Blog Post
    ...recommended for plasmids up to 25 kb, most cloners should be fine, especially using plasmids in the 10-12...Other Plasmid Cloning Techniques like SLIC and Golden Gate Cloning Learn How to Verify Your Plasmid Read...Reference Pages Choose Your Perfect Plasmid Backbone Find Plasmids for Your Research ...made genomic SDM relatively straightforward, but plasmid-based SDM has lagged behind. While commercial kits... REPLACR (Recombineering of Ends of Linearized Plasmids After PCR). Current site directed mutagenesis...mutation, then use those to PCR amplify the template plasmid. The parental, nonmutated DNA is digested with ...of background colonies carrying the unmodified plasmid. For insertions, it’s possible to use the same ...

  12. New CRISPR Web Resources and #12DaysofCRISPR Recap

    Type
    Blog Post
    ...introduce you to the plasmid class covered on that page. To help you find popular plasmids, we’ve also added...changes we’ve made: New CRISPR plasmid pages We’ve given our CRISPR Plasmids and Resources a fresh new design...design, sorting the plasmids we have available by category. Each plasmid page now has a helpful graphic... new plasmid pages that cover some of the hottest CRISPR tools. Base Edit: Base editing plasmids allow...revolution, Addgene has distributed over 100,000 CRISPR plasmids. But that’s not our only job - we strive to also...up with new developments (and, of course, find plasmids that will be useful to you.) With these goals...mind, we set out to update our web resources and plasmid pages to make them more useful and easy to navigate...

  13. Starter Guide to induced Pluripotent Stem Cells (iPSCs) Part 2:  Reprogramming and Transdifferentiation

    Type
    Blog Post
    ... and A.B. Lassar, Expression of a single transfected cDNA converts fibroblasts to myoblasts. Cell, 1987... [6], plasmid electroporation (episomal) [7, 8] and mRNA transfection [9]. Many of the plasmids used for...cells [18] Red blood cells [19, 20] Osteoclasts and osteoblasts [21, 22] Cardiomyocytes [23, 24] Smooth...treatment of mouse fibroblasts with 5-azacytidine led to their conversion into myoblasts [39]. In 2000, Ferber...Jeon, O.H., et al., Human iPSC-derived osteoblasts and osteoclasts together promote bone regeneration in...can be easily isolated from donors. Apart from fibroblasts, human keratinocytes from hair pluck, peripheral...them into different cell lineages [13]. Over the last decade, many cell types have been successfully differentiated...

  14. In Vivo Biotinylation of Bacterial Fusion Proteins

    Type
    Blog Post
    ...about Popular Plasmids in the Repository Learn about New Plasmid Technologies in our Hot Plasmid Articles ...to create a plasmid based on the biotin ligase BirA backbone (Eric Campeau, Addgene plasmid #26624) was...inserting BirA into a pMal plasmid (NEB) thus creating the pMal-T-Avi-His/BirA plasmid now available through...other Hot Plasmids Find Additional Posts on Mol Bio Techniques Find Additional Posts on Plasmid Technologies...controlled easier than formation of GST dimers. Avi-Tag plasmid for improved biotinylation selectivity A common...exploit the best of both worlds. We generated a plasmid that produces biotin ligase and a protein of interest...and a 6X HisTag for affinity purification (see plasmid map and cartoon below). Including biotin in your...

  15. Tips for a 1st time AAV user (by a Rookie AAV user)

    Type
    Blog Post
    ...packaging plasmids. The cells need three different plasmids to produce AAV: 1) the RepCap plasmid which provides...AAV plasmid standard and AAV transfer plasmids. It’s easy to contaminate a workspace with plasmid, not...going to lie, AAV transfer plasmids are kind of a pain to work with. These plasmids contain ITRs which are...AAV’s protein shell, or capsid; 2) the pHelper plasmid which expresses adenovirus genes which help mediate...mediate AAV viral replication; and 3) the transfer plasmid which encodes a transgene of interest that’s packaged...structures that are prone to deletion from the transfer plasmid. The good news is there’s a few simple ways to ...multiple bacterial cultures of your AAV transfer plasmid at 30 °C instead of 37 °C and then screen for ITR...

  16. Pooled Library Amplifications

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    Blog Post
    ...recombined plasmid to overtake full-size plasmids (or for smaller plasmids to overtake larger plasmids, if the...How many unique plasmids does your library contain?), the variance of size of plasmids in your library...lead to issues such as recombination and loss of plasmid diversity, so Addgene strives to distribute aliquots...library (How important is it that every single plasmid in your library is present, and how will you ensure...30 to 200 times as many colonies as individual plasmids in your library. As an example, if you want 100x... your library, and your library contains 5,000 plasmids, your goal for your transformation would be to...libraries often contain thousands, if not millions, of plasmids, they are generally electroporated into electrocompetent...

  17. The MTA at Addgene

    Type
    Blog Post
    ...organizations choose to share their plasmids under an OpenMTA, which allows plasmids to be used for both research...If you’ve deposited or requested plasmids from Addgene, then you’ve been involved in the material transfer...Even though this process is essential to sharing plasmids around the globe, it can be a bit of a mystery...use agreements. The UBMTA, which is the MTA most plasmids at Addgene are shared under, is well-understood...we are not a party (signatory) to the MTAs for plasmids in our repository. Instead, we facilitate the ...depositor and requestor) organizations. Over the last twenty years, we’ve developed a pretty smooth system...institutes. But they’re not! The vast majority of our plasmids are distributed under the UBMTA, which is widely...

  18. Tips and Tricks for Using Golden Gate Modular Cloning (MoClo)

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    Blog Post
    ...blog Plasmids 101: Modular Cloning Plasmids 101: Modular Cloning Applications and Kits Plasmids 101: ...through your plasmid stocks. To avoid cross-contamination, we recommend retrieving plasmids from your original...Calculation for the plasmid volume for MoClo experiments. The term ‘insert’ refers to plasmids carrying each...will find some useful tips to help you make your plasmid construction with MoClo a walk in the park.  Before...promoter, a coding sequence, a terminator, or other plasmid part? Do you want to fuse your protein to any tags...restriction enzymes, and it will create the resulting plasmid map and identify any mismatches that are preventing...Level 0, Level 1, and Level 2 assemblies using the plasmid sequence files provided with every Addgene MoClo...

  19. Viral Production at Addgene

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    Blog Post
    ...the transfer plasmid, a plasmid encoding rep and serotype-specific cap genes, and a plasmid encoding adenoviral..., and is routinely tested for mycoplasma contamination using mycoplasma detection kits. Cell line is maintained... tested for mycoplasma contamination. To date, Addgene has never had a case of mycoplasma contamination.... coli. In addition, plasmids are typically prepared using endotoxin-free plasmid purification kits.   ...contamination is minimized by using an endotoxin-free plasmid purification protocol. To minimize the immunogenic...normalization purposes are being used. Quality Control Mycoplasma Our 293T cell line was obtained directly from...offline and discarded. Confirmation of the transfer plasmid Addgene uses a rigorous barcode matching system...

  20. Early Career Researcher Toolbox: Free Online Molecular Biology Tools

    Type
    Blog Post
    ... in one place. Plasmid mapping These tools are for viewing, editing or making plasmid maps, but can also...make plasmid maps. Free for academic users. Serial Cloner: Free desktop-based software for plasmid design...design and mapping. ApE (A plasmid Editor): A free, donation-based plasmid analysis tool including editing...Utah.   Figure 1: A plasmid map created in SnapGene of Blue Flame plasmid pET21a-BirA.   DNA / ...Primer design. Plasmid mapping. DNA sequence analysis. We all have our favorite tools for tackling these...The free SnapGene Viewer is great for looking at plasmid maps and viewing sequencing traces, while the paid... paid version provides more tools for plasmid mapping and design (Figure 1). Benchling: While you might...
Showing: 201 - 220 of 1038 results