PDF pCR -Blunt II- TOPO - Thermo Fisher Scientific 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. I have even tried to increase the insert (2:1 . 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. The Fisher Scientific Encompass Program offers items which are not part of our distribution portfolio. Efficient cleavage requires at least two copies of the NaeI recognition sequence. pCR4Blunt-TOPO (linearized) Linearized vector with bound topoisomerase, for TOPO cloning of blunt PCR products. 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. pCR-Blunt II- K n TOPO in 3519 bp site: 1099-1236 bases gene bases sit ORF:586-888: bases406-425 bases 433-448 1237-2031 Zeocin PDF CRISPR/Cas9-Directed Genome Editing of Cultured Cells - Harvard University & Engineering, Model For Research Use Only. We will not share your information for any other purposes. Sequence Author: Thermo Fisher (Invitrogen), SnapGene is the easiest way to plan, visualize and document your everyday molecular biology procedures. I am trying to clone this insert in PCR BLUNT end TOPO vector. PDF TOPO PCR cloning solutions - Thermo Fisher Scientific I have used 4 ul of salt. This material may be used without restriction by academic, nonprofit, and governmental entities, except that the source must be cited as www.snapgene.com/resources. Plasmid Lox-Stop-Lox TOPO from Dr. Tyler Jacks's lab contains the insert loxP-STOP-loxP and is published in Genes Dev. However, we are committed to improving your shopping experience. Therapeutic homology-independent targeted integration in retina and You may not be able to create an account or request plasmids through this website until you upgrade your browser. Receive the latest news, hot plasmids, discounts and more. Today, Invitrogen TOPO PCR cloning kits are still among the fastest, simplest, and most flexible cloning solutions available. This vector is NOT available from Addgene. How can I track requests for my plasmids? Your feedback has been submitted. PCR-Blunt II-Topo kit) and incubate at 37C with shaking for 1 hr. No ligase, post-PCR procedures, or PCR primers containing specific sequences are required. pCR-Blunt II-TOPO Sequence and Map - SnapGene Exceptional. How can I be notified when a plasmid from a specific lab or paper is available? However, we are committed to improving your shopping experience. Efficient cleavage requires at least two copies of the RsrII recognition sequence. | Download SnapGene Viewer. These products typically do not have pictures or detailed descriptions. Sequence Author: Thermo Fisher (Invitrogen) Open in SnapGene. Learn more . 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 Flexible. Systems, Research pCR-Blunt II-TOPO vector map and sequence - novoprolabs.com TOPO Blunt-End for Subcloning | Thermo Fisher Scientific - CN 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. Learn more, Please note: Your browser does not fully support some of the features used on Addgene's website. Citing NovoPro. Kit includes linearized and topoisomerase I-activated pCR-Blunt II-TOPO Vector for 5-minute benchtop ligations without ligase. Try SnapGene for Free. 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. Spread 100 l of the transformation mixture . Close the tube and incubate for 10 minutes at room temperature. no. What do I need to know about the customs and importation process for my country? 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. Carefully open the tube and add 20 l of sterile water to dissolve the DNA. Efficient cleavage requires at least two copies of the NgoMIV recognition sequence. This page is informational only - this vector is NOT available from Addgene - please contact the manufacturer for further details. Thermo Fisher Scientific, Save time by having your items shipped automatically. Privacy Policy. There is a problem with the plasmid I received. Genome This website uses cookies to ensure you get the best experience. 71013-3) . 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). All contact information provided shall also be maintained in accordance with our 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. The maps, notes, and annotations in the zip file on this page are copyrighted material. Efficient cleavage requires at least two copies of the BpmI recognition sequence. Please note: Your browser does not support the features used on Addgene's website. What is virus associated DNA, and why do I have to order it. Learn about the latest plasmid technologies and research tools. PDF TOPO TA Cloning - Molbio Learn about the latest plasmid technologies and research tools. The pCRBlunt II-TOPO vector contains: The Fisher Scientific Encompass Program offers items which are not part of our distribution portfolio. PDF Zero Blunt PCR Cloning Kit - Thermo Fisher Scientific Includes the Zero Blunt TOPO vector containing the ccdB gene for positive selection, only permitting growth of plasmid vectors with recombinants. PDF QUICK REFERENCE Zero Blunt TOPO PCR Cloning Kit - Fisher Sci How It Works The plasmid vector (pCRII-TOPO or pCR2.1-TOPO) is supplied linearized with: Single 3 thymidine (T) overhangs for TA Cloning The pCRBlunt II-TOPO vector contains: The Fisher Scientific Encompass Program offers items which are not part of our distribution portfolio. 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? Sequence Author: Thermo Fisher (Invitrogen) Open in SnapGene. All contact information provided shall also be maintained in accordance with our no. PCR-Blunt II-Topo kit (Invitrogen, cat. Receive the latest news, hot plasmids, discounts and more. If you run into any problems registering, depositing, or ordering please contact us at [emailprotected] Receive the latest news, hot plasmids, discounts and more. 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. How do I place an order? We will not share your information for any other purposes. 3. Offers the fastest and easiest method for high-efficiency (cloning of blunt-end PCR products amplified with proofreading thermostable polymerases. 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. Kit includes linearized and topoisomerase I-activated pCR-Blunt II-TOPO Vector for 5-minute benchtop ligations without ligase. Please use the form below to provide feedback related to the content on this product. How do I prepare and deposit my plasmids? All contact information provided shall also be maintained in accordance with our Explore Over 2.7k Plasmids: I.M.A.G.E. Please sign in to view account pricing and product availability. Addgene: Vector Database - pCR blunt II TOPO Can anyone provide help with this TOPO Blunt cloning? Zero Blunt TOPO PCR Cloning Kit, without competent cells Backbone Vector backbone pCR-Blunt II-TOPO (Search Vector Database) Backbone manufacturer Invitrogen Vector type Mammalian Expression, CRISPR Growth in Bacteria Bacterial Resistance (s) Kanamycin, 50 g/mL Growth Temperature 37C Fields, Pathways Learn about the latest plasmid technologies and research tools. 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. Analyze Sequence: pCR blunt II TOPO. Search Try SnapGene for Free. Please use the form below to provide feedback related to the content on this product. Zero Blunt PCR Cloning Kit - Thermo Fisher Scientific Eco R I sites flanking the PCR product insertion site for easy excision of inserts. 11. pCR4-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. Addgene - Analyze Sequence 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, Invitrogen Zero Blunt TOPO PCR Cloning Kit, without competent cells Welcome to Vector Database! Sign Up for Our Newsletter. Fisher Scientific is always working to improve our content for you. Have questions about your order, deposit, or a plasmid? Not for use in diagnostic procedures. 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. Addgene: Lox-Stop-Lox TOPO Thermo Fisher Scientific. Consortium Plasmids Source/Author: Invitrogen (Life Technologies) Selection Marker: Neomycin/G418 (Geneticin) Copy Number: Zero Blunt TOPO PCR Cloning Kit, with pCR-Blunt II-TOPO Vector, One Not for use in diagnostic procedures. The kits include linearized and topoisomerase I-activated pCR-Blunt II-TOPO Vector for 5-minute benchtop ligations without ligase (1). 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. Zero Blunt TOPO PCR Cloning Kit (Cat. Please use the form below to provide feedback related to the content on this product. Genome 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. Please note: Your browser does not support the features used on Addgene's website. I have used 1 ul of 10 ng/ul insert and 1 ul of TOPO vector. How do I place an order? 2. Has anyone grown pCR-BluntII Topo in-house? | ResearchGate 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. Les kits de clonage par PCR Zero Blunt contiennent: un vecteur PCR-Blunt linaris, une ligase dADN T4 ExpressLink, 5X tampons de ligase dADN T4 ExpressLink, un modle de contrle, des dNTPs, de leau strile et des amorces directes et inverses M13. 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. Download Plasmid. Search Two vials of lyophilized plasmid will pCR-Blunt II-TOPO (linearized) Sequence and Map - SnapGene Subscribe. Does Addgene accept orders by fax, phone or email? Fields, Pathways Zero Blunt TOPO PCR Cloning Kit supplied with the PureLink Quick Plasmid Miniprep Kit (Cat. Not for use in diagnostic procedures. SnapGene Viewer is free software that allows molecular biologists to create, browse, and share richly annotated sequence files. Fisher Scientific is always working to improve our content for you. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. How can I be notified when a plasmid from a specific lab or paper is available? Please sign in to view account pricing and product availability. Not for use in diagnostic procedures. 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. Price Information Two vials of lyophilized plasmid will be delivered, each vial is about 5g. 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. 15(24):3243-8. . Vector database is a digital collection of vector backbones assembled from publications and commercially available sources. 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). Have questions about your order, deposit, or a plasmid? By continuing to use this site, you agree to the use of cookies. Consortium Plasmids | More Plasmid Sets. Not for use in diagnostic procedures. These products typically do not have pictures or detailed descriptions. We appreciate your feedback. 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. By clicking Submit, you acknowledge that you may be contacted by Fisher Scientific in regards to the feedback you have provided in this form. Commercial entities must contact GSL Biotech LLC for permission and terms of use. Have questions about your order, deposit, or a plasmid? Consortium Plasmids|More Plasmid Sets No matches HomePlasmidsI.M.A.G.E. Invitrogen Backbone size w/o insert . The quality of . 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. 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. Consortium PlasmidspCR-Blunt II-TOPO What strain of bacteria does my stab contain? Can be used to generate nested deletions for sequencing. Medium, and a supercoiled control plasmid. Editing, Cloning How can I track requests for my plasmids? 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 contains: EcoRI sites flanking the PCR product insertion site for easy excision of inserts Backbone manufacturer. Thermo Fisher Scientific. PDF Zero Blunt TOPO PCR Cloning Kit - Thermo Fisher Scientific How do I prepare and deposit my plasmids? 1333 Xinlong Road, Minhang District, Shanghai, China, Zip-code: 201101, Copyright 2014 NovoPro Bioscience Inc. All rights reserved. For transformation of electrocompetent cells, a diagram of the multiple cloning site, and a manual, see www.lifetechnologies.com/support or contact Technical Support. We appreciate your feedback. 2001 Dec 15. Contact Addgene. 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. By continuing to use this site, you agree to the use of cookies. Bioz Stars score: 86/100, based on 1 PubMed citations. Systems, Research Pcr Blunt Ii Topo Backbone Vector | Thermo Fisher | Bioz 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). Search Room 229, Building 26, No. Simple. Sign Up. 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. ("TOPO" Cloning") for the direct insertion of Taq polymerase-amplified PCR products into a plasmid vector. Resistant to Kanamycin (KanR) and Zeocin (ZeoR). The pCRBlunt II-TOPO vector is designed to clone blunt-ended PCR products generated by thermostable proofreading polymerases such as Platinum Pfx DNA Polymerase. Invitrogen Zero Blunt TOPO PCR Cloning Kit, with pCR -Blunt II-TOPO 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. |, Publications no. & Engineering, Model Fisher Scientific is always working to improve our content for you. Does Addgene accept orders by fax, phone or email? & ORFs. 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. 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.
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