The pCRBlunt II-TOPO vector used in this kit comes with lethal ccdB gene for efficient selection of blunt-ended PCR recombinants products generated by thermostable proofreading polymerases. Efficient cleavage requires at least two copies of the NgoMIV recognition sequence. Learn more, Please note: Your browser does not fully support some of the features used on Addgene's website. Learn more, Download our file to copy and paste plasmid data, Learn more about Addgene materials from user-contributed reports describing AAV and antibody experiments, Basic analysis for a user-entered sequence; includes restriction sites and map, Digital collection of empty plasmid backbones from publications and commercially available sources. Exceptional. |, Publications How can I be notified when a plasmid from a specific lab or paper is available? The Zero Blunt TOPO PCR Cloning Kit for Subcloning, without competent cells allows the flexibility of using your own strain for increased flexibility so competent cells and S.O.C. Welcome to Vector Database! The Zero Blunt PCR Cloning Kit offers an easy method for high-efficiency (>80%) cloning of blunt-end PCR products amplified with proof-reading, thermostable DNA polymerases. These products typically do not have pictures or detailed descriptions. pCRBlunt II-TOPO vector: designed for proofreading polymerasesThe pCRBlunt II-TOPO vector is designed to clone blunt-ended PCR products generated by thermostable proofreading polymerases such as Platinum Pfx DNA Polymerase. The standard in cloning When it comes to cloning, TOPO cloning technology has been a reliable partner for thousands of scientists for over ten years. This is a free resource for the scientific community that is compiled by Addgene. Blunt DNA Cloning Vectors, TOPO Cloning Vectors, pCR, Electrophoresis, Western Blotting and ELISA, Chromatography and Mass Spectrometry Reagents, Laboratory Syringe Needles and Accessories, Lab Coats, Aprons, and Other Safety Apparel, Sharps Disposal Containers and Accessories, Classroom Laboratory Supplies and Consumables, Applied Biosystems TaqMan Assay and Arrays Search Tool, Applied Biosystems TaqMan Custom Assay Design Tools, Applied Biosystems Custom qPCR Primers and TaqMan Probes Tool, Chemical Storage and Management Resource Center, EcoRI sites flanking the PCR product insertion site for easy excision of inserts, Kanamycin and Zeocin resistance genes for your choice of selection in, SP6 promoter/primer site for in vitro RNA transcription and sequencing, M13 forward and reverse primer sites for sequencing or PCR screening, Fast and easy: go from PCR to clones in just three steps and in as little as 5 minutes hands-on time, Efficient: obtain up to 95% clones with correct insert, Proven: reliable performance for over a decade with over 4,000 citations, Simple: no ligase, post-PCR procedures, or PCR primers containing specific sequences are required, Vector: pCRBlunt II-TOPO vector: contains the, Cloning method: TOPO cloning: Topoisomerase Ibased, 5-minute ligation of blunt-ended PCR products amplified with proofreading thermostable polymerases, Competent cells: Various options: choose from kits with either general, high-efficiency, bacteriophage T1-resistant, fast-growing competent cells (available separately) or use your own, Antibiotic resistance: Ampicillin (AmpR), Kanamycin (KanR). These products typically do not have pictures or detailed descriptions. Explore Over 2.7k Plasmids: I.M.A.G.E. This is a free resource for the scientific community that is compiled by Addgene. What do I need to know about the customs and importation process for my country? The pCRBlunt II-TOPO vector used in this kit comes with lethal ccdB gene for efficient selection of blunt-ended PCR recombinants products generated by thermostable proofreading polymerases. Eco R I sites flanking the PCR product insertion site for easy excision of inserts. The quality of . Sequence Author: Thermo Fisher (Invitrogen) Open in SnapGene. Les kits cellulaires comptents contiennent des cellules d, Copyright 2006-2023 Thermo Fisher Scientific Inc. All rights reserved, Colonnes et cartouches de chromatographie, Consommables en plastique et fournitures de laboratoire, Afficher toutes les catgories de produits, Spectroscopie, analyse lmentaire et isotopique, Voir toutes les applications et techniques, Services aux organisations de dveloppement et de fabrication sous contrat (CDMO) et pour les essais cliniques, Consultez toutes les rubriques d'aide et d'assistance. Le kit de clonage de PCR Zero Blunt permet de cloner facilement et efficacement (>80%) les produits de PCR aux extrmits franches, qui ont t amplifis par des ADN polymrases thermostables activit correctrice. PCR-Blunt II-Topo kit) and incubate at 37C with shaking for 1 hr. ("TOPO" Cloning") for the direct insertion of Taq polymerase-amplified PCR products into a plasmid vector. Sequence Author: Thermo Fisher (Invitrogen) Open in SnapGene Try SnapGene for Free Download Plasmid|Download SnapGene Viewer Explore Over 2.7kPlasmids:I.M.A.G.E. pCR-Blunt II-TOPO Parental vector for TOPO cloning of blunt PCR products. Can be used to generate nested deletions for sequencing. How do I place an order? pCR-Blunt II-TOPO (linearized) Linearized vector for TOPO cloning of blunt PCR products. Does Addgene accept orders by fax, phone or email? Please use the form below to provide feedback related to the content on this product. However, we are committed to improving your shopping experience. Zero Blunt TOPO PCR Cloning Kit supplied with the PureLink Quick Plasmid Miniprep Kit (Cat. Introduction Product Overview Description of the system The Zero Blunt PCR Cloning Kit is designed to clone blunt PCR fragments (or any blunt DNA fragment) with a low background of nonrecombinants. 3. We appreciate your feedback. We appreciate your feedback. | Download SnapGene Viewer. Consortium Plasmids|More Plasmid Sets No matches HomePlasmidsI.M.A.G.E. & Engineering, Model Citing NovoPro. Speed is less than 5000g. The pCRBlunt II-TOPO vector contains: The Fisher Scientific Encompass Program offers items which are not part of our distribution portfolio. For Research Use Only. & Engineering, Model Learn more, Download our file to copy and paste plasmid data, Learn more about Addgene materials from user-contributed reports describing AAV and antibody experiments, Basic analysis for a user-entered sequence; includes restriction sites and map, Digital collection of empty plasmid backbones from publications and commercially available sources. This website uses cookies to ensure you get the best experience. The - pCR-Blunt vector contains the lethal E. coli ccdB gene fused to the C-terminus of LacZ (Bernard et al., 1994).Ligation of a blunt PCR fragment disrupts expression Today, Invitrogen TOPO PCR cloning kits are still among the fastest, simplest, and most flexible cloning solutions available. Vector database is a digital collection of vector backbones assembled from publications and commercially available sources. Thermo Fisher Scientific. Have questions about your order, deposit, or a plasmid? How can I track requests for my plasmids? Fisher Scientific is always working to improve our content for you. Spread 100 l of the transformation mixture . pCRBlunt II-TOPO vector: designed for proofreading polymerasesThe pCRBlunt II-TOPO vector is designed to clone blunt-ended PCR products generated by thermostable proofreading polymerases such as Platinum Pfx DNA Polymerase. Not for use in diagnostic procedures. For transformation of electrocompetent cells, a diagram of the multiple cloning site, and a manual, see www.lifetechnologies.com/support or contact Technical Support. Sequence Author: Thermo Fisher (Invitrogen) Open in SnapGene. The standard in cloning When it comes to cloning, TOPO cloning technology has been a reliable partner for thousands of scientists for over ten years. Kit includes linearized and topoisomerase I-activated pCR-Blunt II-TOPO Vector for 5-minute benchtop ligations without ligase. PCR-Blunt II-Topo kit (Invitrogen, cat. ( How to cite ) Sequence Information Sequences (4) Ordering This material is available to academics and nonprofits only. Sequence Author: Thermo Fisher (Invitrogen), SnapGene is the easiest way to plan, visualize and document your everyday molecular biology procedures. no.K2800-02) is shipped with an additional box containing reagents for plasmid purification (Box 3). Editing, Cloning Zero Blunt TOPO PCR Cloning Kits offers the fastest and easiest method for high-efficiency (cloning of blunt-end PCR products amplified with proofreading thermostable polymerases. Subscribe. The kits include linearized and topoisomerase I-activated pCR-Blunt II-TOPO Vector for 5-minute benchtop ligations without ligase (1). Search The maps, notes, and annotations in the zip file on this page are copyrighted material. M13 forward and reverse primer sites for sequencing or . Resistant to Kanamycin (KanR) and Zeocin (ZeoR). Basic Vector Information Vector Name: pCR-Blunt II-TOPO Antibiotic Resistance: Kanamycin Length: 3519 bp Type: I.M.A.G.E. Each kit has the Zero Blunt TOPO vector (see figure) containing the ccdB gene for positive selection, only permitting growth of plasmid vectors with recombinants so you can obtain up to 95% clones with correct insert. Room 229, Building 26, No. Efficient cleavage requires at least two copies of the NaeI recognition sequence. Download Plasmid. By clicking Submit, you acknowledge that you may be contacted by Fisher Scientific in regards to the feedback you have provided in this form. 1333 Xinlong Road, Minhang District, Shanghai, China, Zip-code: 201101, Copyright 2014 NovoPro Bioscience Inc. All rights reserved. How can I track requests for my plasmids? Genome In addition, a Zero Blunt TOPO Cloning Kit is available combined with a Invitrogen PureLink Quick Plasmid Miniprep Kit (50 preps) for fast plasmid purification of your TOPO-cloned inserts for downstream analysis. Your feedback has been submitted. Commercial entities must contact GSL Biotech LLC for permission and terms of use. The pCR Blunt II-TOPO vector features: Contains the ccdB gene for positive selection; EcoRI sites flanking the PCR product insertion site for easy excision of inserts; Kanamycin and Invitrogen Zeocin resistance genes for your choice of selection in E. coli; SP6 promoter/primer site for in vitro RNA transcription and sequencing; M13 forward and reverse primer sites for sequencing or PCR screening Please note: Your browser does not support the features used on Addgene's website. Close the tube and incubate for 10 minutes at room temperature. We appreciate your feedback. What is an MTA/Who is authorized to sign? Each Zero Blunt TOPO for subcloning Kit contains linearized and topoisomerase I-activated pCR Blunt II-TOPOVector, Salt Solution, dNTPs, Control Template and Primers, M13 Forward and Reverse Primers, One Shot Chemically Competent or ElectrocompE. coli, S.O.C. These products typically do not have pictures or detailed descriptions. Invitrogen Backbone size w/o insert . How can I be notified when a plasmid from a specific lab or paper is available? Receive the latest news, hot plasmids, discounts and more. Medium, and a supercoiled control plasmid. Not for use in diagnostic procedures. Bioz Stars score: 86/100, based on 1 PubMed citations. Receive the latest news, hot plasmids, discounts and more. 450245) is shipped with only the Zero Blunt TOPO PCR Cloning reagents (Box 1). Consortium Plasmids Source/Author: Invitrogen (Life Technologies) Selection Marker: Neomycin/G418 (Geneticin) Copy Number: The pCR-Blunt II-TOPO Vector also features: For Research Use Only. The PCR fragment was subcloned in reverse complementary orientation in PCR-Blunt II-TOPO (Invitrogen) before cloning in a pAAV2.1 backbone by In-Fusion (Takara) using EcoRI restriction sites. 11. Includes the Zero Blunt TOPO vector containing the ccdB gene for positive selection, only permitting growth of plasmid vectors with recombinants. Systems, Research All contact information provided shall also be maintained in accordance with our Consortium PlasmidspCR-Blunt II-TOPO By clicking Submit, you acknowledge that you may be contacted by Fisher Scientific in regards to the feedback you have provided in this form. pCR-Blunt II-TOPO . & ORFs. Plasmid Lox-Stop-Lox TOPO from Dr. Tyler Jacks's lab contains the insert loxP-STOP-loxP and is published in Genes Dev. We will not share your information for any other purposes. Privacy Policy. Please use the form below to provide feedback related to the content on this product. Carefully open the tube and add 20 l of sterile water to dissolve the DNA. Privacy Policy. Thermo Fisher Scientific, Save time by having your items shipped automatically. Can be used to generate nested deletions for sequencing. The Fisher Scientific Encompass Program offers items which are not part of our distribution portfolio. Zero Blunt TOPO PCR Cloning Kit (Cat. Le kit de clonage de PCR Zero Blunt sappuie sur le vecteur de clonage multifonctions PCR-Blunt et la ligase dADN ExpressLinkT4 de faon gnrer temprature ambiante et en cinq minutes un produit de ligature. Learn more, Please note: Your browser does not fully support some of the features used on Addgene's website. 2. Search The pCRBlunt II-TOPO vector is designed to clone blunt-ended PCR products generated by thermostable proofreading polymerases such as Platinum Pfx DNA Polymerase. How do I prepare and deposit my plasmids? Fast accurate construct design for all major molecular cloning techniques, Validate sequenced constructs using powerful alignment tools, Customize plasmid maps with flexible annotation and visualization controls, Automatically generate a rich graphical history of every edit and procedure, Gain unparalleled visibility of your plasmids, DNA and protein sequences, Annotate features on your plasmids using the curated feature database, Store, search, and share your sequences, files and maps. SP6 promoter/primer site for in vitro RNA transcription and sequencing. Includes the Zero Blunt TOPO vector containing the ccdB gene for positive selection, only permitting growth of plasmid vectors with recombinants. What do I need to know about the customs and importation process for my country? Fisher Scientific is always working to improve our content for you. no. Learn more . Get the cloning results you can count on Twenty years ago, Invitrogen TOPO technology became known for superior quality and game-changing innovation in cloning. Parental sequencing vector for TOPO TA cloning of PCR products. Flexible. Analyze Sequence: pCR blunt II TOPO. Electrophoresis, Western Blotting and ELISA, Chromatography and Mass Spectrometry Reagents, Laboratory Syringe Needles and Accessories, Lab Coats, Aprons, and Other Safety Apparel, Sharps Disposal Containers and Accessories, Classroom Laboratory Supplies and Consumables, Applied Biosystems TaqMan Assay and Arrays Search Tool, Applied Biosystems TaqMan Custom Assay Design Tools, Applied Biosystems Custom qPCR Primers and TaqMan Probes Tool, Chemical Storage and Management Resource Center, EcoRI sites flanking the PCR product insertion site for easy excision of inserts, Kanamycin and Zeocin resistance genes for your choice of selection in, SP6 promoter/primer site for in vitro RNA transcription and sequencing, M13 forward and reverse primer sites for sequencing or PCR screening, Fast and easy: go from PCR to clones in just three steps and in as little as 5 minutes hands-on time, Efficient: obtain up to 95% clones with correct insert, Proven: reliable performance for over a decade with over 4,000 citations, Simple: no ligase, post-PCR procedures, or PCR primers containing specific sequences are required, Vector: pCRBlunt II-TOPO vector: contains the, Cloning method: TOPO cloning: Topoisomerase Ibased, 5-minute ligation of blunt-ended PCR products amplified with proofreading thermostable polymerases, Competent cells: Various options: choose from kits with either general, high-efficiency, bacteriophage T1-resistant, fast-growing competent cells (available separately) or use your own, Antibiotic resistance: Ampicillin (AmpR), Kanamycin (KanR). The kits include linearized and topoisomerase I-activated pCR-Blunt II-TOPO Vector for 5-minute benchtop ligations without ligase (1). The pCRBlunt II-TOPO vector is designed to clone blunt-ended PCR products generated by thermostable proofreading polymerases such as Platinum Pfx DNA Polymerase. Press Release Team Advisors Partners Contact Bioz Stars Bioz vStars 86 Buy from Supplier Structured Review Thermo Fisher pcr blunt ii topo backbone vector Pcr Blunt Ii Topo Backbone Vector, supplied by Thermo Fisher, used in various techniques. What is virus associated DNA, and why do I have to order it. Molecular Genetics TOPO Vector Cloning Vector Human Molecular Genetics Most recent answer Magnus Wohlfahrt Rasmussen University of Copenhagen 11th Apr, 2015 That's true, you can make a regular. By continuing to use this site, you agree to the use of cookies. Sequence Author: Thermo Fisher (Invitrogen) Open in SnapGene Try SnapGene for Free Download Plasmid|Download SnapGene Viewer Explore Over 2.7kPlasmids:TOPO Cloning Vectors|More Plasmid Sets No matches Sign Up for Our Newsletter. Vector database is a digital collection of vector backbones assembled from publications and commercially available sources. no. How It Works The plasmid vector (pCRII-TOPO or pCR2.1-TOPO) is supplied linearized with: Single 3 thymidine (T) overhangs for TA Cloning Have questions about your order, deposit, or a plasmid? Try SnapGene for Free. Copyright 2006-2023 Thermo Fisher Scientific Inc. All rights reserved, Sulen und Kartuschen fr die Chromatographie, Kunststoffartikel und Zubehr fr das Labor, Spektroskopie, Element- und Isotopenanalyse, Alle Themen fr Hilfe und Support anzeigen, Status und Nachverfolgung von Bestellungen, Zero Blunt TOPO PCR Cloning Kit for Subcloning, without competent cells, EcoRI sites flanking the PCR product insertion site for easy excision of inserts, Kanamycin and Invitrogen Zeocin resistance genes for your choice of selection in, M13 forward and reverse primer sites for sequencing or PCR screening, Flexibility to use your own competent cell strain, Electrocompetent cells for highest transformation efficiency, Includes PureLink Quick Plasmid Miniprep Kit. The pCRBlunt II-TOPO vector contains: EcoRI sites flanking the PCR product insertion site for easy excision of inserts Learn about the latest plasmid technologies and research tools. How do I place an order? The pCR Blunt II-TOPO vector is designed to clone blunt-ended PCR products generated by thermostable proofreading polymerases such as Platinum SuperFi DNA Polymerase. Two vials of lyophilized plasmid will Search Offers the fastest and easiest method for high-efficiency (cloning of blunt-end PCR products amplified with proofreading thermostable polymerases. If you run into any problems registering, depositing, or ordering please contact us at [emailprotected] Fisher Scientific is always working to improve our content for you. This vector is NOT available from Addgene. pCR-Blunt II-TOPO 3519 bp 500 1000 1500 2000 2500 3000 3500 BsaXI, AseI, +25 +4 KasI SphI NcoI, +3 +9 +1 +3 +5 +1 AlwNI BssSI, +1 DrdI, AflIII, Copyright 2006-2023 Thermo Fisher Scientific Inc. All rights reserved, Zero Blunt TOPO PCR Cloning Kit, with pCR-Blunt II-TOPO Vector, One Shot TOP10 Chemically Competent, Spectroscopy, Elemental and Isotope Analysis. USER GUIDE Zero BluntTOPO PCR Cloning Kit For 5-minute cloning of blunt-end PCR products Catalog Numbers K2800-20, K2800-40, K2800-J10, K2820-20, K2820-40, K2830-20, K2860-20, K2860-40, K2800-02, 450245 Revision C.0 Publication Number MAN0000062 For Research Use Only. Resistant to Kanamycin (KanR) and Zeocin (ZeoR). You may not be able to create an account or request plasmids through this website until you upgrade your browser. By clicking Submit, you acknowledge that you may be contacted by Fisher Scientific in regards to the feedback you have provided in this form. Please sign in to view account pricing and product availability. Simple. K2800-20) including One Shot Top10 Chemically Competent E. coli cells (other competent cells for cloning may also be used) Sterilized glass beads (EMD Millipore, cat. Please note: Your browser does not support the features used on Addgene's website. PCR products were cloned into PCR-Blunt II-TOPO (Invitrogen) and single clones were sequenced to confirm the identity of the PCR products before NGS analysis. What is virus associated DNA, and why do I have to order it. The pCRBlunt II-TOPO vector contains: The Fisher Scientific Encompass Program offers items which are not part of our distribution portfolio. no. Not for use in diagnostic procedures. Medium are not provided. Thermo Fisher Scientific, Zero Blunt TOPO PCR Cloning Kit provides a highly efficient, 5-minute, one-step cloning strategy ("TOPO Cloning") for the direct insertion of blunt-ended PCR products amplified with proofreading thermostable polymerases into a plasmid vector. Receive the latest news, hot plasmids, discounts and more. Backbone manufacturer. Each kit has the Zero Blunt TOPO vector (see figure) containing the, Blunt DNA Cloning Vectors, TOPO Cloning Vectors, pCR, Copyright 2006-2023 Thermo Fisher Scientific Inc. All rights reserved, Zero Blunt TOPO PCR Cloning Kit, without competent cells, Spectroscopy, Elemental and Isotope Analysis. Electrophoresis, Western Blotting and ELISA, Chromatography and Mass Spectrometry Reagents, Laboratory Syringe Needles and Accessories, Lab Coats, Aprons, and Other Safety Apparel, Sharps Disposal Containers and Accessories, Classroom Laboratory Supplies and Consumables, Applied Biosystems TaqMan Assay and Arrays Search Tool, Applied Biosystems TaqMan Custom Assay Design Tools, Applied Biosystems Custom qPCR Primers and TaqMan Probes Tool, Chemical Storage and Management Resource Center, Kanamycin and Zeocin resistance genes for your choice of selection in, SP6 promoter/primer site for in vitro RNA transcription and sequencing, M13 forward and reverse primer sites for sequencing or PCR screening, Selected Zero Blunt TOPO PCR Cloning Kits are available in combo kits combined with a PureLink Quick Plasmid Miniprep Kit (50 preps) for fast plasmid purification of your TOPO-cloned inserts for downstream analysis. Privacy Policy. Have questions about your order, deposit, or a plasmid? I have even tried to increase the insert (2:1 . pCR4Blunt-TOPO (linearized) Linearized vector with bound topoisomerase, for TOPO cloning of blunt PCR products. Genome Not for use in diagnostic procedures. Fields, Pathways Consortium Plasmids | More Plasmid Sets. Please sign in to view account pricing and product availability. No ligase, post-PCR procedures, or PCR primers containing specific sequences are required. Learn about the latest plasmid technologies and research tools. SnapGene Viewer is free software that allows molecular biologists to create, browse, and share richly annotated sequence files. Introduction Follow these instructions to TOPO Clone your blunt-end PCR product into pCR-Blunt II-TOPO and transform the reaction into chemically competent E. coli cells. All contact information provided shall also be maintained in accordance with our There is a problem with the plasmid I received. For Research Use Only. Editing, Cloning By continuing to use this site, you agree to the use of cookies. For NGS library preparation, 10 ng of DNA from the PCR products were used as input for the NEBNext Ultra II DNA Library Prep Kit for Illumina (New England Biolabs).
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