Protocol/8

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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 12-16<sup>o</sup>C.


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