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FAQ

Home > TECHNICAL > FAQ
How can I increase the efficiency of EZ-Fusion™ Cloning reaction?

A: Use accurate amounts of vector and insert. It is better to use 100 ng of vector if the size is less than 10 kb and 200 ng of the vector if the size is larger than 10 kb. Calculate the amount of insert DNA as following method.

(Insert size) / (vector size) x (amount of vector) x 2


High efficiency could be obtained by purification of the vector and insert DNA. Increase the total reaction volume to 20 µl if the concentration of insert and vector is low. However, to obtain high efficiency use only 5 µl of EZ-Fusion™ cloning reaction for transformation.


Using large volume of EZ-Fusion™ cloning reaction for trasfomation does not increase the efficiency. Enzymes in EZ-Fusion™ cloning reaction are toxic to some competent cells, therefore in common use only 2.5 µl of the EZ-Fusion™ cloning reaction for the transformation.


Proper reaction temperature and time is 37 and 15 minutes.

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