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We narrowed to 24 results for: Gateway entry

Showing: 1 - 20 of 24 results

  1. Plasmids 101: Gateway Cloning

    Type
    Blog Post
    ... the Gateway cloning method and its advantages. An introduction to Gateway technology The Gateway cloning...to generate entry clones Entry vector attL sites for recombination Used to generate entry clones by TOPO...systems. With the appropriate entry and destination vectors, one can use Gateway to clone a gene of interest...1: Generate an Entry Clone There are a few different ways to generate our desired entry clone - human ...program to design Gateway plasmids.    Figure 3: Method A to create an entry clone: recombination... an attL-containing entry vector.   Figure 4: Method B to create an entry clone: TOPO cloning ... an attL-containing entry vector.   Figure 5: Method C to create an entry clone: Restriction cloning...

  2. Quickest Way to Deposit Plasmids: The Deposit Spreadsheet

    Type
    Blog Post
    ...the most involvement with Step 1, and, while data entry can be a chore, we’ve made it easy to send us your...information in a deposit spreadsheet. The data entry process On the Addgene home page, if you scroll ...top of the page, you’ll see two options for data entry: “Submit Online Deposit” and “Download Deposit Spreadsheet...blog post for a thorough guide to our online data entry process. Figure 1: Begin the deposit process by...template vector”. Cloning Method Choose from: Gateway cloning, gene synthesis, Gibson cloning, ligation-independent...For plasmids containing the ccdB gene, such as Gateway vectors, choose the ccdB Survival strain. For plasmids...deposit spreadsheet will help expedite your data entry and make it easier than ever to share your discoveries...

  3. Hot Plasmids - March 2019 - Anti-CRISPR, 2in1 Cloning, Fluorescent Voltage Indicators, and Photoswitchable Proteins

    Type
    Blog Post
    ...Similar to traditional Gateway™ cloning, the GOI first needs to be cloned into an entry vector. The 2in1 cloning...introduced the “2in1” vector cloning system - a Gateway compatible approach to simultaneously clone two... cloning system contains two entry vectors (pUC57-L1L4 and pUC57-L3L2) that offer the advantage of using...NC, CN, and CC). The 4 kits also contain the two entry vectors pUC-L1L4 and pUC-L3L2. Find the 2in1 plasmid... the GOIs into the 2in1 vector instead of the GatewayTM cloning BP reaction, where insertion of the GOI...

  4. Plasmids 101: CcdB - The Toxic Key to Efficient Cloning

    Type
    Blog Post
    ...recently developed Gibson Assembly Cloning and Gateway® cloning technologies, as well as several others... to thoroughly screen colonies for the insert. Gateway® technology (developed by InvitrogenTM) is essentially... a separate post. Focusing on the ccdB aspect, Gateway takes advantage of the same principle that cells...http://teachline.ls.huji.ac.il/72682/tutorials/gateway/introduction.html demonstrating the principal behind...and suggestions, and even to submit a guest blog entry of your own! References 1. Bernard, P. “Positive...Additional Web Resources OpenWetWare's CcdB Page Gateway Cloning Page at Techline ...focus on a unique gene that makes the popular GatewayTM method possible: ccdB. But what is ccdB, what ...

  5. Plasmids 101: Golden Gate Cloning

    Type
    Blog Post
    ...ligated into a vector containing those overhangs. Entry DNA overhangs may be present in the original plasmid... 30-minute reaction. The destination vector and entry vector(s) are placed in a single tube containing...can be assembled with Golden Gate. The popular Gateway cloning system produces constructs with an attB...want to consider using an alternative method like Gateway cloning or Gibson assembly. Another important consideration...

  6. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ...systems. The MultiSite Gateway Kit contains a set of 12 promoter-less Gateway® entry clones to be used with...on the Multisite Gateway® technology from Life Technologies. The MultiSite Gateway Kit is a customizable...available as a standard entry clone. ORFs from an ORFeome Gateway library or any PCR-amplified ORF cloned into... phage integrase system which is the basis for Gateway cloning. The combination of all 11 att pairs in...customizable Gateway®-based toolkit that allows scientists to generate fusion proteins from any ORF available...heterologous promoter outside of the Multisite Gateway® cassette, you can specifically express your new...with other entry clones encoding the ORFs of interest: Buj et al., BMC Molecular Biology. 2013 August...

  7. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    ... with Multisite Gateway cloning to generate the final construct. Up to eight entry vectors can be used...component plasmids from building blocks via Golden GATEway Cloning Frustrated by the complexity of assembling...combining the advantages of Golden Gate and Multisite Gateway cloning methods developed by the lab of Joachim...just the set of tools that you need. The Golden GATEway cloning kit simplifies the cloning process for ...a desired component are cloned into Golden Gate entry vectors via traditional restriction enzyme cloning...cloning, or annealing of oligonucleotides. These entry vectors are used in a Golden Gate cloning step to... used for each Multisite Gateway compatible destination vector for a maximum of 24 elements assembled ...

  8. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    ...Although only wt hCas9 entry vectors are supplied with the kit, you can clone your own entry vectors containing...repression), and Fok1-dCas9 (dimeric nuclease). Gateway assembly method Frew Lab Multiple Lentiviral Expression...expressing wt humanized Cas9 and up to three gRNAs. Entry vectors containing the U6 promoter and the gRNA ...should be compatible with type IIS enzyme BfuAI. Gateway cloning is then used to combine the multiple gRNAs...

  9. How to Deposit Your Plasmids with Addgene

    Type
    Blog Post
    ...the top right of each data entry page. Once you have completed the data entry process for all of the plasmids...will be saved and the page will open a new set of entry fields to describe the next insert. Repeat as needed...For plasmids containing the ccdB gene, such as Gateway vectors, we recommend the ccdB Survival strain....the "Submit Deposit" button to complete the data entry. Clicking the "Submit Deposit" button will allow...

  11. Empty Backbones - Choosing Your Perfect Plasmid Backbone

    Type
    Collection
    ...Ubiquitin pYPQ203 (pMDC32-Ubi1) - Multisite Gateway T-DNA entry plasmid (attR1 & attR2); Zea mays Ubi1 promoter...-ccdB - Yeast Gateway expression vector (see more Gateway vectors) pCAP03... TRP1 Yeast pGBKCg - Gateway compatible Y2H vector LEU2 Yeast pGADT7-GW - Gateway compatible Y2H vector...vector - AD pGADCg - Gateway compatible Y2H vector BASTA Plant pGTQL1211YN - Gateway compatible BiFC vector...myc-nuc - pEntry vector that adds a C-terminal nuclear localization signal pGADCg - Gateway yeast expression...collection page Yeast GAL4, PGK, ADH1, ADE2, TRP1 Gateway destination vectors - Inducible... proteins, and more pGBKT7-GW - Gateway compatible Y2H vector - BD pRPR1_gRNA_handle_RPR1t...

  13. Anatomy of a Plasmid Page at Addgene

    Type
    Blog Post
    ..., so some of our older plasmids may not have an entry in this field. If you’ve deposited a plasmid without...may require strains such as ccdB survival (for Gateway plasmids).  We also like to store plasmids that...

  14. Fluorescent Protein Guide: Empty Backbones

    Type
    Collection
    ...pMH-Halo tag - Mammalian Gateway Destination Vector pENTR4-HaloTag (w876-1) - Gateway Entry Vector pCS2+/C-Halo...Mammalian Expression pENTR4-SNAPf (w878-1) - Gateway Entry Vector pCS2+/C-SNAPf , pCS2+/N-SNAPf - Xenopus...Mammalian Expression pENTR4-CLIPf (w877-2) - Gateway Entry Vector pET51b-His-TEV-CLIP-tag - Bacterial Expression...

  15. CRISPR References and Information

    Type
    Collection
    ...; dCas9 activator gateway donors: pAC84 , pAC1 , pAC147 , pAC148 , pAC149 ; Gateway destination: pAC90...encyclopedia of Class 2 CRISPR systems with wiki entries describing enzyme activity, experimental considerations...

  16. CRISPR Plasmids - Empty gRNA Vectors

    Type
    Collection
    ...encyclopedia of Class 2 CRISPR systems with wiki entries describing enzyme activity, experimental considerations...Mammalian BbsI yes, activate S. pyogenes Jaenisch piCRg Entry 58904 Mammalian BbsI yes, cut S. pyogenes Huangfu...assembly. Gersbach MuLE System Mammalian MultiSite Gateway recombination-based cloning of complex polycistronic...plasmids available for use in this Golden Gate and Gateway based assembly method: plasmids for assembly of...a single T-DNA binary vector of choice through Gateway recombination. Qi Multiplex Conditional Mutagenesis...

  17. Tetracycline Inducible Expression

    Type
    Collection
    ...promoter rtTA3 Wei Xu 26803 pEnt L1L3 EF1a-tTA-2 Gateway entry vector to express tTA from EF1α promoter, for...Tet-On vector for inducible expression; insert with Gateway cloning and puromycin selection rtTA-Advanced Tight...

  18. Molecular Cloning Techniques

    Type
    Guide
    ...completed via Gateway cloning. Addgene's collection contains thousands of Gateway donor , Gateway entry , and ...backbone. Created with BioRender.com. Gateway Recombination Cloning Gateway cloning is a recombination-based...via a proprietary BP clonase, creating an entry clone. The entry clone now has recombined attL sites flanking...made an entry vector containing your DNA of interest (or obtained one of the thousands of entry vectors...experimental goal. Although Gateway cloning is a simple and efficient cloning method, Gateway vectors and recombination...conditions. Read more in our Gateway Cloning blog post . Figure 2: Summary of Gateway cloning. Created with ...further reducing ligation efficiency. TOPO-ready Gateway entry plasmids are also available, allowing for rapid...

  19. Gamma-Retroviral Vector Guide

    Type
    Guide
    ...including restriction enzyme, Gibson Assembly, or Gateway. Some transfer plasmids may have limited restriction...compatible with certain cloning methods (such as a Gateway destination vector), so be sure to confirm your...yield of DNA. However, if the plasmid contains a Gateway cassette containing the ccdB gene, a ccdB-resistant...Needle sticks and ripped gloves are common points of entry. Biosafety should always be considered with respect...packaging cell line). Infection The natural process of entry and multiplication of a pathogen, like viruses or...the viral particles is removed and stored or centrifuged to concentrate. Crude or concentrated virus can...gamma-retroviral vector purification by high-speed centrifugation for CAR T cell manufacturing. Molecular Therapy...

  20. Adeno-associated virus (AAV) Guide

    Type
    Guide
    ...including restriction enzyme, Gibson Assembly, or Gateway. Some transfer plasmids may have limited restriction...compatible with certain cloning methods (such as a Gateway destination vector), so be sure to confirm your...yield of DNA. However, if the plasmid contains a Gateway cassette containing the ccdB gene, a ccdB-resistant...Needle sticks and ripped gloves are common points of entry. Biosafety should always be considered with respect...Blood-brain barrier (BBB) Layer of cells that prevent the entry of toxins and pathogens from the blood to the CNS...packaging cell line). Infection The natural process of entry and multiplication of a pathogen, like viruses or...Cells AAV Purification by Iodixanol Gradient Ultracentrifugation AAV Titration by ddPCR AAV Titration by qPCR...
Showing: 1 - 20 of 24 results