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en:over-hang [2013/06/09 19:10] (current)
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 +<select Language>​
 +Protruding region of dsDNA or the sequence of the protruding region.
 +|{{bio:​overhanging.jpg?​100|}}\\ [[http://​​wiki/​Wspinaczka|Image]]|{{bio:​overhang.png|}}|
 +=====Biological application=====
 +If over-hang of termini of a cloning vector and a insert are complimentary,​ they hybridizes and are [[Ligase|ligated]] effectively. The direction of the insert can be controlled by using two differenct over-hang sequences.
 +=====How to produce=====
 +  *Digest with a restriction enzyme.
 +  *Add a recognition sequence of a restriction enzyme to 5'​-terminus of primers used in PCR
 +  *Use an over-hang fragment produced with Taq PCR.  TdT activity of Taq adds an [[Nucleobase#​purine|adenine]] to end of the fragment ([[TA-cloning]])
 +  *Anneal ssDNA (or ssRNA).
 +  *[[Ligase]]
 +  *[[PCR]]
 +  *[[TA-cloning]]
 +  *[[TdT activity]]