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Invitrogen™ Zero Blunt™ PCR Cloning Kit, without competent cells
Description
Includes
Zero Blunt™ PCR cloning kits contains: linearized pCR™-Blunt vector, ExpressLink™ T4 DNA ligase , 5X ExpressLink™ T4 DNA ligase buffer, control template, dNTPs, sterile water, and M13 forward and reverse primers. Competent cell kits contain One Shot™ chemically competent E. coli, S.O.C. medium, and a supercoiled control plasmid.
The Zero Blunt™ PCR Cloning Kit uses the multipurpose cloning vector pCR™-Blunt and ExpressLink™ T4 DNA Ligase to generate a ligation product in a five-minute, room-temperature ligation step.
The pCR-Blunt vector provides:
- EcoR I sites flanking the PCR product insertion site for excision of inserts
- T7 promoter/primer site for in vitro RNA transcription and sequencing
- M13 forward and reverse primer sites for sequencing or PCR screening
The Zero Blunt PCR Cloning Kit is designed to clone blunt PCR fragments (or any blunt DNA fragment) with a low background of non-recombinants. The pCR-Blunt vector contains the lethal E. coliccdB gene fused to the C-terminus of LacZα (Bernard et al., 1994). Ligation of a blunt PCR fragment disrupts expression of the lacZα-ccdB gene fusion permitting growth of only positive recombinants upon transformation. Cells that contain non-recombinant vector are killed when the transformation mixture is plated.
Kit Configurations
The Zero Blunt PCR Cloning Kit is offered in a variety of configurations: with One Shot™ TOPO10 Chemically Competent E. coli (K2700-20 and K2700-40) and without competent cells (K2750-20 and K2750-40) in 20- and 40- reaction kit sizes.
- Fast and convenient: 5-minute, room-temperature ligation
- Efficient: ccdB gene for positive selection results in >80% clones with correct insert
- Flexible: choice of kanamycin or Zeocin™ resistance for flexible antibiotic selection
Cloning, PCR Cloning
Order Info
Shipping Condition: Dry Ice
Specifications
Specifications
| Bacterial or Yeast Strain | Not Included |
| Cloning Method | Blunt PCR |
| Content And Storage | Zero Blunt™ PCR cloning kits contain linearized pCR™-Blunt vector, ExpressLink™ T4 DNA ligase , 5X ExpressLink™ T4 DNA ligase buffer, control template, dNTPs, sterile water, and M13 forward and reverse primers. Store all components at -20°C. All reagents are guaranteed stable for 6 months when properly stored. |
| Format | Kit |
| Product Line | TOPO, Zero Blunt |
| Product Type | PCR Cloning Kit |
| Promoter | T7 |
| Quantity | 20 Reactions |
| Vector | Blunt DNA Cloning Vectors |
Safety and Handling
For Research Use Only. Not for use in diagnostic procedures.
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