The pENTR™/D-TOPO® Cloning Kits utilize a highly efficient, 5-minute cloning strategy Ω("TOPO® Cloning") to directionally clone a blunt-e♣nd PCR product into a vector for entry into the Gateway® ™System or the MultiSite Gateway® System. Blunt-end PCR products clone directionally at gεreater than 90% efficiency, with no ligase, post-PCR procedu&res, or restriction enzymes required.

The kit comes with everything necessary to clone and select your PCR ₽amplified gene of interest:

• Gateway® System Ready—Rapidly shuttle cloned genes between multiple vector systems
• Fast and Easy—Go from PCR to Gateway® Entry clone in just 3 steps an≠d as little as ~5 minutes hands on time
• Efficient—Achieve over 90% clones with correct insert in the right di↓rection
• Proven—Reliable performance for over a decade

pENTR™ /D-TOPO® Cloning Kits Overview

Simple Access to the Gateway® System
For access to the Gateway® System, just PCR amplify your gene ofφ interest and add the product straight to the provided topoisomerase charged pENTR$™/D-TOPO® vector, incubate 5 minutes, and transform the provided competent E. coli cells. The resulting attL containing Gateway® Entry clones are÷ ready for efficient recombination with your choice of G∞ateway® Destination vectors.

Optimized pENTR™/D-TOPO® Vector
The pENTR™/D-TOPO® vector (Figure 1) includes M13 and T7 primer sequencing sites and δattL recombination sites flanking the PCR product insertion site. This all₹ows clones to be easily sequence verified and recoδmbined into your choice of attR containing Gateway® destin<ation vectors. A Kanamycin resistance gene and a pUC origin are used for sele×ction and high-copy propagation in E. coli.

Simplified Directional Cloning
With Directional TOPO® cloning technology there is no need for PCR clean up, vector prepar"ation, or other time-intensive DNA manipulation steps. Just add your PCR reaction stra©ight to the provided topoisomerase charged vector, incubate 5 m✘inutes, transform, and obtain up to 90% directionally inserted clon<es. A four base overhang on the vector pairs with a four base sequence designed into the forwarσd primer used in your PCR reaction to provide directio×nality to the topoisomerase ligation reaction (Fig←ure 2).

The Power of Gateway® Recombination Cloning Technoσlogy
Gateway® recombination cloning technology circumvents the limitations of restriction mediated φcloning, enabling you to access virtually any expression system  in a simple one hour, 99%-efficient and reversible, Gateway→® recombination reaction. The ability to move the ÷same sequence of DNA between different vectors without using restriction enzymes, ligase, subclonβing steps, screening of countless colonies, or re-sequencing will help save you time, money, andΩ effort.

Leading Cloning Technologies
When it comes to cloning, TOPO® cloning technology and Gateway®≤ recombination cloning technology have been a reliable partner for thousands of s‌cientists for over ten years. Fast, simple to use, and efficient, TOPO® cloning and Ga>teway® recombination allow for rapid cloning and subsequent tranσsferring of genes between a wide assortment of Gateway® expression vec↕tors.

詳細說(shuō)明(míng)

常用(yòng)規格

內(nèi)容和(hé)儲存

pENTR™/D-TOPO® Cloning Kit contains pENTR/D-TOPO® vector, dNTPs, salt≥ solution, sterile water, and universal M13 sequencing primers.

Store components at -20°C.