Invitrogen™ Zero Blunt™ TOPO™ PCR Cloning Kit, without competent cells

Supplier:  Invitrogen™ 450245

Catalog No. 45-024-5

Includes: Linearized and topoisomerase I-activated pCR™Blunt II-TOPO™ vector; Salt solution; dNTPs; Control template; M13 forward and reverse primers; Sterile water

Description

Includes the Zero Blunt TOPO vector containing the ccdB gene for positive selection, only permitting growth of plasmid vectors with recombinants. Resistant to Kanamycin (KanR) and Zeocin™ (ZeoR).
pCRBlunt II-TOPO vector: designed for proofreading polymerases
The pCRBlunt II-TOPO vector is designed to clone blunt-ended PCR products generated by thermostable proofreading polymerases such as Platinum™ Pfx DNA Polymerase. The pCRBlunt 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 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

pCRBlunt II-TOPO clone selection
pCRBlunt II-TOPO allows for direct selection of recombinants via disruption of the lethal E. coli gene, ccdB. The vector contains the ccdB 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 cloning
Using 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 transform E. coli competent cells.
Efficient cloning
With up to 95% of clones carrying the desired insert, you can screen fewer clones, saving you time and money. 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.
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. 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 options
The 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. Nos. K2820-20, K2820-40)
Fast growth: Mach1™-T1R Chemically Competent E. Coli (Cat. No. K2830-20)
Provide your own: for flexibility and to save money (Cat. No. 450254)
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 is also available.

• 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: pCR™Blunt II-TOPO vector: contains the ccdB gene, allowing for direct selection of recombinants
• Cloning method: TOPO cloning: Topoisomerase I–based, 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)

Cloning, PCR Cloning

Order Info

Shipping Condition: Dry Ice

Specifications

• Bacterial or Yeast Strain: Not Included
• Cloning Method: Blunt TOPO™
• Format: Kit
• Product Type: PCR Cloning Kit
• Quantity: 25 Reactions
• Vector: Blunt DNA Cloning Vectors, TOPO Cloning Vectors, pCR

• Antibiotic Resistance Bacterial: Kanamycin (KanR), Zeocin™ (ZeoR)
• Content And Storage: Box 1:
  • Linearized and topoisomerase I-activated pCR™Blunt II-TOPO™ vector
  • Salt solution
  • dNTPs
  • Control template
  • M13 forward and reverse primers
  • Sterile water
  
  Store at -5 to -30°C.
  All reagents are stable for 6 months when properly stored.
• Product Line: TOPO™, Zero Blunt™
• Promoter: SP6
• Shipping Condition: Dry Ice

For Research Use Only. Not for use in diagnostic procedures.