Bio-rad Ligation and Transformation Module Manuale Utente Scaricare il pdf (Pagina 20)

One advantage to sticky-end ligation is that it makes directional

cloning possible. If it is desirable to have the insert in one orientation

only (for instance, in the A Æ B direction in the vector, but not in the

B Æ A direction), then the insert and vector can both be digested with two

different restriction enzymes so that their ends are asymmetric. This is important

if the DNA insert is a cDNA encoding a protein to be expressed in the

transformed cell. When this is done, only the complementary ends will ligate

and the insert will have a single orientation in the ligation products.

Blunt end digestion.

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Bio-rad Ligation and Transformation Module Manuale Utente Pagina 20