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en:blunting_sticky_ends [2013/06/09 10:10] (current)
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 +======Blunting sticky ends======
 +Sticky-ends can be blunted with appropriate enzymes
 +
 +Following enzymatic activities are utilized for blunting of DNA ends.
 +  *5'​->​3'​ polymerase activity to '​fill-in'​.
 +  *Deletion by exonuclease activity.
 +  *Digestion by endonuclease.
 +Notice that the structure of the ends depends on the activities of enzyme.
 +
 +=====DNA polymerase=====
 +Almost commercially available blunting kits contain the DNA polymerase. ​ It fills-in 3'​-sticky ends and deletes 5'​-sticky ends.  The blunting kits are useful because you don't have to prepare the reaction buffer containing dNTP.
 +
 +{{bio:​t4polymerase_product.png|}}
 +
 +
 +=====S1 Nuclease=====
 +S1 Nuclease is one of endonuclease that recognize single-strand DNA or RNA.  It can cut phosphodiester bondings where the site is not hybridized. ​ Therefore, it can blunt DNA ends as well as remove loop structures on the dsDNA.
 +
 +{{bio:​s1nuclease_product.png|}}
 +
 +Images were created by [[http://​wiki.symplus.co.jp/​cmaster:​en:​Blunting%20sticky%20ends|ConstructMaster]].
 +
 +
 +======Enzymes of kits======
 +^Vendor^Enzyme in the blunting kit^
 +|TaKaRa Bio|T4 DNA Polymerase|
 +|Nippon gene|T4 DNA Polymerase|
 +|TOYOBO|KOD Polymerase|
 +(on 2006.3.10)
 +
 +
 +======Links======
 +
 +  *[[T4_DNA_Polymerase|T4 DNA polymerase]]
 +
 +
 +{{tag>​blunt polymerase nuclease}}