Invitrogen™ Zero Blunt™ PCR Cloning Kit

Offers an easy method for high-efficiency (>80%) cloning of blunt-end PCR products amplified with proofreading, thermostable DNA polymerases

Overview
Brand: Invitrogen™

Manufacturer Part Number: K270020

ZERO BLUNT PCR CLONING KIT

UNSPSC: 41106610

Code: 50

Additional Details:
Additional Details: Weight: 0.10000kg



Product Code. 10216382

Quantity Price
1 £ 272.0 / 1 set
EU Stock 30
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For Research Use Only. All usage must comply with product instructions.
 

Description and Specification

Specification

Product Type Kit, PCR Cloning
Bacterial/Yeast Strain TOP10
Includes 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. Competent cell kits contain One Shot chemically competent E. coli, S.O.C. medium, and a supercoiled control plasmid
For Use With (Application) Cloning, PCR Cloning
Quantity 20 reactions

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
How Zero Blunt PCR Cloning Works
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 & 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