Thermo Fisher, K280020SC, Zero Blunt™ TOPO™ PCR Cloning Kit, with One Shot™ TOP10 Chemically Competent E. coli (Supply Center Packaging)

Zero Blunt™ TOPO™ PCR Cloning Kit, with One Shot™ TOP10 Chemically Competent E. coli (Supply Center Packaging) 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. Each kit has the Zero Blunt™ TOPO™ vector (see figure) containing theccdB gene for positive selection, only permitting growth of plasmid vectors with recombinants. Zero™ Blunt™ TOPO™ kits are available with a variety of competent cells, or without competent cells, depending on your needs and budget. This product comes in Supply Center packaging, which features an additional outer box that holds both the TOPO™ vector box and the competent cells box. This special packaging helps ensure that visitors to a Supply Center obtain the complete kit.
Features of the Zero™ Blunt™ TOPO™ PCR Cloning Kits:Fast and easy—go from PCR to clones in just 3 steps and in as little as 5 minutes hands-on timeEfficient—obtain up to 95% clones with correct insertProven—reliable performance for over a decade with over 4,000 citationsSimple —no ligase, post-PCR procedures, or PCR primers containing specific sequences are required.
Zero Blunt™ TOPO™ Cloning Kits—overviewVector:pCR™Blunt II-TOPO™ vector—contains theccdB gene, allowing for direct selection of recombinantsCloning method:TOPO™ cloning—Topoisomerase I–based, 5-minute ligation of blunt-ended PCR products amplified with proofreading thermostable polymerasesCompetent cells:Various options—choose from kits with either general, high-efficiency, bacteriophage T1-resistant, fast-growing competent cells, or use your own.
pCR™Blunt II-TOPO™ vector—designed for proofreading polymerasesThe pCR™Blunt II-TOPO™ vector is designed to clone blunt-ended PCR products generated by thermostable proofreading polymerases such as Platinum™PfxDNA Polymerase. The pCR™Blunt II-TOPO™ vector contains:
• EcoRI sites flanking the PCR product insertion site for easy excision of inserts
• Kanamycin and Zeocin™ resistance genes for your choice of selection inE. coli
• SP6 promoter/primer site forin vitroRNA transcription and sequencing
• M13 forward and reverse primer sites for sequencing or PCR screeningpCR™Blunt II-TOPO™ clone selectionpCR™-Blunt II-TOPO™ allows for direct selection of recombinants via disruption of the lethalE. coligene,ccdB. The vector contains theccdB gene fused to the C-terminus of the LacZα fragment. Ligation of a blunt-end PCR product disrupts expression of the LacZα-ccdB gene fusion, permitting growth of only positive recombinants upon transformation. Cells that contain non-recombinant vector are killed upon plating. Therefore, blue/white screening is not required.
Simplified TOPO™-based cloningUsing TOPO™ cloning technology, there is no need for PCR primers containing specific sequences, post-PCR procedures, vector preparation, or other time-intensive DNA manipulation steps. Just add your PCR reaction straight to the provided topoisomerase-charged vector, incubate 5 minutes, and transformE. colicompetent cells.
Efficient cloningWith up to 95% of clones carrying the desired insert, you can screen less clones, saving you time and money. The pCR™Blunt II-TOPO™ vector used in this kit comes with lethalccdB gene for efficient selection of blunt-ended PCR recombinants products generated bythermostable proofreading polymerases.
The standard in cloningWhen it comes to cloning, TOPO™ cloning technology has been a reliable partner for thousands of scientists for over ten years. Fast, simple-to-use, and efficient, TOPO™ cloning has been applied to many different vectors for a wide array of applications.
Zero Blunt™ TOPO™ PCR Cloning™ Kit optionsThe Zero Blunt™ TOPO™ PCR Cloning Kit for direct insertion of blunt-ended PCR products into a plasmid vector can be purchased with a variety of competent cells that deliver different advantages depending upon your needs:General cloning: TOP10 cells (Cat. No. K2800-20, K2800-40)High-efficiency cloning: TOP10 Electrocomp™ Cells (Cat. No. K2860-01, K2860-40)General cloning, bacteriophage T1 resistance: DH5α-T1R (Cat. No. K2820-20, K2820-40)Fast growth: Mach1™-T1R Chemically CompetentE. Coli(Cat. No. K2830-20)Provide your own: for flexibility and to save money (Cat. No. 450254)We also offer a version of the kit that includes a PureLink™ Quick Plasmid Miniprep Kit (Cat. No. K2800-02) for use in isolation of clean, sequencing-ready, recombinant plasmid.
Bacterial or Yeast Strain: TOP10
Cell Type: Chemically Competent
Cloning Method: Blunt TOPO™
Product Line: One Shot™
Product Type: PCR Cloning Kit
Promoter: SP6
Quantity: 25 Reactions
Shipping Condition: Dry Ice
Vector: Blunt DNA Cloning Vectors
Format: Kit
Unit Size: Each