Protocol/8

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Protocol 8: Ligation
 
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Reagents:
 
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* Vector
 
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* Insert
 
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* MilliQ buffer
 
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* 5X ligase buffer
 
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* T4 DNA ligase
 
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** [http://tools.invitrogen.com/content/sfs/manuals/t4dnaligase_5U_man.pdf]
 
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Procedure:
 
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*Volumes of vector and insert may vary, but a typical 20 ul reaction might be:
 
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{|
 
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| MilliQ water || 11 ul
 
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|-
 
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| 5x ligase buffer || 4 ul
 
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|-
 
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| Vector (200 ng/ul) || 1 ul
 
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|-
 
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| Insert (200 ng/ul) || 3 ul
 
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|-
 
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| Ligase || 1 ul
 
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|}
 
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*Incubate at R/T for 1 hours or overnight at 12-16<sup>o</sup>C.
 
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[[Team:Alberta/Notebook/protocols| Back]]
 
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Latest revision as of 17:15, 26 October 2010