Protocol/8

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*Volumes of vector and insert may vary, but a typical 20 ul reaction might be:
*Volumes of vector and insert may vary, but a typical 20 ul reaction might be:
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| MilliQ water || 11 ul
| MilliQ water || 11 ul
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| Ligase || 1 ul
| Ligase || 1 ul
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*Incubate at R/T for 1 hours or overnight at 12-16<sup>o</sup>C.
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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]]
[[Team:Alberta/Notebook/protocols| Back]]
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{{Team:Alberta/endMainContent}}

Revision as of 01:23, 26 October 2010

TEAM ALBERTA

Protocol 8: Ligation


Reagents:

  • Vector
  • Insert
  • MilliQ buffer
  • 5X ligase buffer
  • T4 DNA ligase
    • [http://tools.invitrogen.com/content/sfs/manuals/t4dnaligase_5U_man.pdf]


Procedure:

  • Volumes of vector and insert may vary, but a typical 20 ul reaction might be:
MilliQ water 11 ul
5x ligase buffer 4 ul
Vector (200 ng/ul) 1 ul
Insert (200 ng/ul) 3 ul
Ligase 1 ul
  • Incubate at R/T for 1 hours or overnight at 12-16oC.


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