Invitrogen™ Zero Blunt™ TOPO™ PCR Cloning Kit, with One Shot™ TOP10 Chemically Competent E. coli cells

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

Includes: Linearized and topoisomerase I-activated pCR™Blunt II-TOPO™ vector; Salt solution; dNTPs; Control template; M13 forward and reverse primers; Sterile water; One Shot™ Chemically Competent or Electrocomp™ E. coli; S.O.C. medium; Supercoiled pUC19 control plasmid

Catalog Number	Mfr. No.	Quantity
K280020	Invitrogen™ K280020	25 Reactions
K280040	Invitrogen™ K280040	50 Reactions
K2800J10	Invitrogen™ K2800J10	10 Reactions

Description

• Each kit has the Zero Blunt™ TOPO™ vector containing the ccdB gene for positive selection, only permitting growth of plasmid vectors with recombinants
• Available with a variety of competent cells, or without competent cells
• Fast and easy—go from PCR to clones in just 3 steps, including a 5-minute cloning reaction
• Efficient—obtain up to 95% clones with correct insert
• Proven—enabling reliable performance for over a decade with over 20,000 citations
• Simple —no ligase, post-PCR procedures, or PCR primers containing specific sequences required
• Accurate—for use with high fidelity DNA polymerases, no need for A-overhang on PCR product
• The pCR™Blunt II-TOPO™ vector is designed for cloning of blunt-ended PCR products generated by thermostable proofreading polymerases such as Platinum™ Pfx DNA Polymerase.
• 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.
• 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.
• Zero Blunt™ TOPO™ PCR Cloning kits 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

Order Info

Shipping Condition: Dry Ice

Specifications

• Bacterial or Yeast Strain: TOP10
• Cloning Method: Blunt TOPO™
• Format: Kit
• Promoter: SP6

• Cell Type: Chemically Competent
• 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.
  
  Box 2:
  • One Shot™ Chemically Competent or Electrocomp™ E. coli
  • S.O.C. medium
  • Supercoiled pUC19 control plasmid
  
  Store at -68 to -85°C.
• Product Type: PCR Cloning Kit

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