5-Minute cloning and positive selection with Zero Blunt TOPO
Invitrogen Zero Blunt TOPO technology provides a highly efficient, 5-minute, one-step cloning strategy 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 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.
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.
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
Performance and value
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 Electrocomp E. coli, S.O.C. Medium, and a supercoiled control plasmid. 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. Medium are not provided.
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.
Zero Blunt TOPO PCR Cloning Kits for subclobing (with pCR Blunt II TOPO vector)
|Includes One Shot Competent E. coli||Advantage||Rxn number||Cat. No.|
|MAX Efficiency DH5 αT1 Phage Resistant (genotype)||
|TOP10 Invitrogen Electrocomp(genotype)||
|Invitrogen Mach1 T1R (genotype)||
|MAX Efficiency DH5 α T1 Phage Resistant (genotype)||
For Research Use Only. Not for use in diagnostic procedures.