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We narrowed to 993 results for: Des

Showing: 81 - 100 of 993 results

  1. Antibodies 101: Multiplex Immunofluorescence

    Type
    Blog Post
    ... Brewer, M., McDonough, L., Zhu, Y., Neumann, E., De Caestecker, M., Gutierrez, D., & Spraggins, J. (2022...between the targets. Or, it might be sufficient to describe or classify the labeled structures visually; for...will vary depending on your specific experimental design. Like selecting an antibody, some of the requirements...

  2. Plasmids 101: Repressible Promoters

    Type
    Blog Post
    ...2022). https://doi.org/10.1038/s41598-022-07852-7 De Boer, Herman A., Lisa K. Comstock & Mark Vasser. ...to the promoter and turning transcription OFF. Despite their bacterial origins, Tet systems function well...promoters can be used in the repressible manner described above, as well as the inducible manner detailed...30 years since the original Tet-Off system was described by Gossen and Bujard, both the promoter and transactivator...cells, and can be used in the repressible manner described above, as well as the inducible manner detailed...tissue- or developmental stage-specific promoter and design a UAS reporter construct, the reporter will only...simple answer is that sometimes it is necessary or desirable to control or look at multiple genes at the same...

  3. Recombinase-based State Machines Enable Order-dependent Logic in vivo

    Type
    Blog Post
    ...Green, Alexander A., et al. "Toehold switches: de-novo-designed regulators of gene expression." Cell 159.4...also be used to design primers to read states by qPCR or Sanger sequencing as described in the Material...hammerhead ribozyme parts are described in Nielsen et al (3) and Lou et al (4). Despite choosing proper register...machine design space is highly degenerate and often times there are multiple register designs that may...recognition sites from the register. If the RSM is designed in such a way that each possible order of inputs...segments in each state of a register enables the design of primer pairs that PCR amplify in some states...registers within a sample using a metric that we describe in the Materials and Methods section of our paper...

  4. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    ...bacterial mutagenesis or gene targeting Victor de Lorenzo's lab has engineered a modular mini-Tn5 vector...on channelrhodopsins – Chronos & Chrimson Through de novo sequencing of 127 algal transcriptomes, as well...components into a destination vector in a predefined order. Finally, the destination vectors are used ...engineering approach, they designed Opto-RTKs, which activate signaling cascades on exposure to low-intensity...years of distributing his Designer Receptors Exclusively Activated by Designer Drugs (DREADD) plasmids ...minimizing confounding secondary effects? An approach described as “knocksideways” (a British idiom for “taking...recombination elements such as FRT or Lox sites, and provides a modular system for easier exchange and re-use...

  5. Luciferase Plasmid Collection

    Type
    Collection
    ...LuxSit-i and neoLux1.2 : Artificial luciferases desined de novo using deep-learning approaches. Split Luciferase...Structurally destabilized firefly luciferases : A set of three increasingly destabilized mutants which...mammalian systems. ID Plasmid Luciferase Type(s) Description PI 64784 pGL3-Basic-IRES Firefly Insertion of...plasmids. ID Plasmid Luciferase Type Promoter Description PI 60226 AAV:ITR-U6-sgRNA(backbone)-pEFS-Rluc...your regulatory element of interest. ID Plasmid Description Gene/Insert PI Return to Top Other Highlighted...

  6. Plasmids 101: Dimers and Multimers

    Type
    Blog Post
    ...Strausbaugh LD, Gray P, Vartak NB, Brown M, Talbot D, Berg DE. pBR322-derived multicopy plasmids harboring large...confirmations like linear and nicked plasmids, as described in this article.   Long-read sequencing Addgene...bacterial colonies using one of the detection methods described above can help ensure the isolation of the monomeric...copy number plasmids causes instability: CoIE1 encodes a determinant essential for plasmid monomerization...Carnes AE, Hodgson CP. Plasmid DNA vaccine vector design: impact on efficacy, safety and upstream production...

  7. CRISPR History and Development for Genome Engineering

    Type
    Collection
    ...X, Scott DA, Mikkelson T, Heckl D, Ebert BL, Root DE, Doench JG, Zhang F. 2014. Genome-Scale CRISPR-Cas9...also like... CRISPR Guide CRISPR Protocols gRNA Design Tools CRISPR Blog Posts The CRISPR revolution shows... store a record of invading phages but also to destroy the phages upon re-exposure. More specifically,...complex binds to the foreign DNA and cleaves it to destroy the invader ( Figure 1 ). Figure 1: An overview...in vitro. In 2013, Cong et al. and Mali et al. described CRISPR-based genome editing in mammalian cell ... of new applications. The first CRISPR papers described two main categories of genome edits. First, the...very low off-target activity due to rationally designed mutations. Researchers have expanded the CRISPR...

  8. Zhang Lab CRISPR Page

    Type
    Collection
    ..., Scott DA, Mikkelsen TS, Heckl D, Ebert BL, Root DE, Doench JG, Zhang F. Science . 2014 Jan 3;343(6166...oligos (design is indicated below) can be cloned into the CRISPR array. The oligos are designed based on...also like... CRISPR Guide CRISPR Protocols gRNA Design Tools CRISPR Blog Posts Jump to: SpCas9 Plasmids...digested using BbsI, and a pair of annealed oligos (design is indicated below) can be cloned scarlessly into...vector before the sgRNA scaffold. The oligos are designed based on the target site sequence (20bp) and need...using SAM: Addgene plasmids #61422 - #61424 were designed for transient transfection : 61422 : Expresses... references are below. Available plasmids are described below: 61591 : PX601; CMV-driven SaCas9; U6-driven...

  9. An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency

    Type
    Blog Post
    ...https://doi.org/10.1093/nar/gku289 Chen C, Fenk LA, de Bono M (2013) Efficient genome editing in Caenorhabditis... DNA double-strand breaks up to 54 bp from the desired insertion site result in efficient editing. Repair...available through Addgene. In 2019, Farboud et al. described another similar co-conversion marker (repair of...whether it be by improved gRNA or repair template design, delivery, screening, and more. Here are some highlights...highlights from the past few years:  Repair template design The Meyer lab meticulously characterized several...of editing. First, they devised guidelines for designing single-stranded repair templates (Farboud et al...Tools such as SapTrap Builder greatly simplifies design of these constructs, allowing novice researchers...

  10. With an Eye Towards the Future, We Look Back at the March for Science

    Type
    Blog Post
    ...policies that “muzzle” government scientists to the de-funding of crucial environmental research programs...and minority communities, where these issues hit hardest. According to the Guttmacher Institute, most women...was supremely proud of Nashville that morning. Despite the rain, about 4,000 supporters of science showed... an agenda that halts current data collection, destroys existing data, and guts future funding.I marched...science belongs to the taxpayers who fund it, and we deserve to know the truth about our world. Jonathan Aguilar...

  11. Plasmids for Stem Cell Research

    Type
    Collection
    ... cell reprogramming factors and wait for cells to de-differentiate. However it may be difficult to decide...blog post Delivery Methods for Generating iPSCs provides an overview of several different reprogramming...find more information. Delivery Method Species Description Article PI Lentivirus Human Expression of human...

  13. Delivery Methods for Generating iPSCs

    Type
    Blog Post
    ... cell reprogramming factors and wait for cells to de-differentiate. However it’s difficult, especially... with peptides that help mediate their transduction, such as the polyarginine peptide described in Zhou...reprogrammer, to decide which method to use. This post provides a brief overview of reprogramming methods with... Paramyxoviridae family of viruses, which also includes measles and mumps. Sendai transduces a wide range...in practice integration can occur. Okita et al describes a protocol for generating iPSCs by plasmid transfection...

  14. Plan Your Experiment

    Type
    Collection
    ...multi-plasmid SAM system Activate Design and Clone Your gRNA Achieving your desired edit comes down to location...on proper gRNA design. While there are some cases where it makes sense to manually design a gRNA, in most...most cases gRNA design software is better and more efficient at designing potential gRNAs for you to choose...protocol. Read more about how to design your gRNA . Synthesize and Clone Desired gRNAs Once your target sequences...selected, it’s time to design your gRNA oligos and clone them into your desired vector. In many cases,...experiment, including selecting the edit type, designing and cloning a gRNA, delivering Cas and the gRNA... Science Guides CRISPR Guide Plan Your Experiment Plan Your CRISPR Experiment...

  15. CRISPR References and Information

    Type
    Collection
    ...CRISPR/Cas design (Link opens in a new window) Program for designing optimal gRNAs. Provides feedback on...window) This tool helps design (10 different prediction scores), clone (primer design), and evaluate gRNAs...Addgene Church gRNA design and cloning gRNA cloning vector PDF, 118 KB Church gRNA design and cloning for ...window) CRISPR/Cas9 reagent generation; gRNA design; HDR design; Deep sequencing Cas9: pMLM3613 dsDNA donor...CRISPR resources and references, including gRNA design tools, CRISPR FAQs, and community forums. CRISPR...protoSpaceJAM is an all-around platform for CRISPR knock-in design at scale. It uses an open-source algorithm to automate... automate and optimize gRNA and HDR donor design. The current version supports SpCas9, SpCas9-VQR, and...

  16. TALEN Plasmids and Kits

    Type
    Collection
    ...Sangamo)-Destination Pawel Pelczar pCAG-T7-TALEN(Sangamo)-Destination constructs were designed for optimal...pC-GoldyTALEN and RCIsricpt-GoldyTALEN are designed as destination vectors for the Voytas lab Golden Gate ...Plasmids pTAL5-BB and pTAL6-BB are designed as alternative destination vectors for the Golden Gate TALEN...TALEN(Sangamo)-FokI-KKR-Destination 40132 pCAG-T7-TALEN(Sangamo)-FokI-ELD-Destination Golden Gate TALEN 2.0...Mendenhall The TALE plasmid vectors described on this page are designed to target and edit the epigenome ...with custom repeat arrays. TALEN Kits TALEN Kit Description Depositing PI FusX TALEN Assembly System Contains... pre-assembled trimer and tetramer RVD domains designed for rapid TALEN assembly. Kiran Musunuru and Chad...

  17. Empty Backbones - Choosing Your Perfect Plasmid Backbone

    Type
    Collection
    ...CMV GFP DEST - Lentiviral Gateway destination vector for gene expression pLenti CMV/TO Zeo DEST - Tet-inducible...lentiviral vector for ORF expression pDest-565 - Gateway destination vector for bacterial expression; His...Membrane localization pWZL-Neo-Myr-Flag-DEST - pDest vector that adds an N-terminal myristoylation...marker pLenti CMV/TO GFP-Zeo DEST (719-1) - 3rd gen lentiviral Gateway destination vector, expression, CMV/...pLenti CMV/TO Zeo DEST - Tet-inducible lentiviral Gateway destination vector ...expression pLenti CMV GFP DEST - Lentiviral Gateway destination vector for gene ...needed for replication in the host cell and are designed for a user to insert a gene of interest. They ...

  18. Easi-CRISPR: Generating Knock-In and Conditional Mouse Models

    Type
    Blog Post
    ...using in mouse transgenics. Designing an Easi-CRISPR knock-in allele To design a knock-in allele at a given... PAM or guide mutation. Designing an Easi-CRISPR conditional allele The design for a floxed conditional...successfully used guides targeting the same or complementary strands, as well as guides facing towards or...How to design a conditional allele with Easi-CRISPR. As in the knock-in example, you’ll design your ...with your desired insertion feature (e.g., a fluorescent protein), you can amplify the desired region using... Figure 5: How to design Easi-CRISPR repair templates using Addgene plasmids. Design PCR primers to add...arduous. First, you’d construct a plasmid with your desired changes, long homology arms and a selection marker...

  19. 27 Hot Plasmids from 2016

    Type
    Blog Post
    ...relaxed gRNA design parameters. To demonstrate the effectiveness of their library design, Moffat laboratory...-strand DNA (ssDNA) oligonucleotides (oligos), and thereby rapidly create desired allele combinations ... pCAG-T7-TALEN(Sangamo)-Destination series, pCAG-Golden-Gate-Esp3I-Destination Takashi Yamamoto pcDNA-TAL-NC2...vector, and besides being used for restoring strain prototrophy, can also aid in designing self-establishing...assay. In their recent publication, Kroeze et al. describe their enhanced TANGO arrestin recruitment assay...optimized, expression validated GPCR sequences to any desired backbone.  Kroeze, et al. Nat Struct Mol Biol....plasmids with the Cross-disciplinary Integration of Design Automation Research lab Moduclar Cloning (CIDAR...

  20. Plasmids 101: Gateway Cloning

    Type
    Blog Post
    ...clone in their desired DNA sequences. Gateway technology relies on the two reactions described below: The ...pLenti CMV Puro DEST (w118-1) or the doxycycline-inducible pLIX_403. The chosen attR destination vector will...between the entry clone and destination vector creates two products: the desires expression clone and a byproduct...produce the desired expression clone. This is possible thanks to the Gateway vectors’ design. They have...vector systems. With the appropriate entry and destination vectors, one can use Gateway to clone a gene ...generated entry clone and the attR sites of the destination vector. This reaction is catalyzed by the LR ...containing the ccdB gene is excised from the destination vector.  Once the BP and/or LR reactions are ...
Showing: 81 - 100 of 993 results