Name: 用限制性酶進行克隆 (Cloning With Restriction Enzymes)
Uploaded: 2021-01-14T05:18:22.000Z
Duration: 1 min 48 s
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Restriction enzymes identify a specific recognition sequence in the DNA, and generate double-stranded

breaks in the DNA duplex. Cleavage results in fragments with either a 5' or 3' overhang,

commonly referred to as sticky ends, or no overhang, referred to as blunt ends.

The 5' phosphates and 3' hydroxyls are maintained after cleavage, leaving the DNA ends ready

to be joined via ligation. The ability of some restriction enzymes to

predictably cleave DNA and generate distinct DNA bands with ligatable ends has made them

an invaluable tool for recombinant DNA technologies. When selecting restriction enzymes for use

in a cloning experiment, it is important to: determine which enzymes will produce ends

compatible with the selected vector confirm that recognition sites do not occur

within the DNA fragment to be cloned and examine the methylation sensitivity

of your selected enzyme to confirm that host methylation by dam and/or dcm methylases will

not block cleavage. NEB offers a variety of online interactive tools to aid in selecting

which enzyme to use, as well as parameters for setting up reactions.

Visit CLONEWITHNEB.com for a full list of products available for this application.