The pCR™8/GW/TOPO® TA Cloning Kit contains the TOPO®-adapted, ready-to-use pCR™8/GW/TOPO® Vector, dNTPs, Salt Solution, Sterile Water, GW Forward and Reverse Sequencing Primers, Control Template and PCR Primers, One Shot® TOP10 Chemically Competent E. coli or One Shot® Mach1™-T1^R Chemically Competent E. coli, S.O.C. Medium, and a pUC19 control plasmid. Store the One Shot® E. coli at -80°C. Store all other components at -20°C. All reagents are guaranteed stable for 6 months when properly stored.

描述

TOPO® TA Cloning® Kits are designed for cloning PCR products directly from a PCR reaction in just 5 minutes. TOPO® TA Cloning® Kits use a pCR™-TOPO® Vector with covalently bound topoisomerase I for fast cloning and recombinants. The newest TOPO® TA Cloning® Kit comes with a Gateway® (GW) entry vector, pCR™8/GW/TOPO® (Figure 1). This vector has:

• 3´-T overhangs for direct ligation of Taq-amplified PCR products
• att sites for rapid recombination into a variety of Gateway® destination vectors
• Novel GW primer sites, within the att regions, located less than 55 base pairs from the PCR product insertion site for convenient sequencing
• Spectinomycin resistance gene for robust selection in E. coli
• EcoR I sites flanking the PCR product insertion site for easy excision of inserts

Selected TOPO® TA Cloning® Kits for entry into the Gateway® Technology are available in combo kits combined with a PureLink™ Quick Plasmid Miniprep Kit (50 preps) for fast plasmid purification of your TOPO®-cloned inserts for downstream analysis.

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