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en:ligase [2013/06/09 10:10] (current)
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 +<select Language>​
 +Ligase connect a 5'​-terminus and 3'​-terminus of a polynucleotide with ATP. This enzyme plays roles in DNA replication and reparing in vivo.
 +\\ ligase+[[ATP]] →
 +===DNA replication===
 +  *Connect 5'​-terminus and 3'​-terminus of '​Okazaki fragments'​ which is polymerized by a polymerase.
 +===DNA reparing===
 +  *Connect 5'​-terminus and 3'​-terminus after BER or NER reparing.
 +====Purpose of use====
 +The ligase is an essential enzyme for cloning technology. ​ Restriction enzymes are compared to '​scissors',​ ligases are compared to '​glue'​ as well. 
 +===About using Ligation-Kits===
 +Commercially available '​ligation-kits'​ contain specialized buffers optimized with salt concentration and detergent concentration. Moreover, some kits contain additional proteins such as a reparing protein, PprA (Nippon gene).
 +We have to be careful for the buffer before transformation because the buffer might suppress the ligation reaction. Therefore, the solution after ligation reaction should be subjected to Ethanol-precipitation method. Some ligation kit contains a buffer that cancel such adverse effect (TaKaRa).
 +Anyway, we can use kits emphasizing '​High-Efficient'​ in ease, but we should understand why it is efficient.
 +  -[[Restriction-modification system]]
 +  -[[ATP]]
 +  -[[ADP]]