USTC/Notebook/DNA ligation

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== DNA Ligation ==

DNA ligases are used with restriction enzymes to insert genes into plasmids.


Materials
*'''DNA plasmids''' double digested previously(eg.XbaI and PstI); . Measure concentration in Nanodrop beforehand. 
*'''Biobricks''' double digested previously(eg.SpeI and PstI )
*equal amount of  T4 ligase.



Procedure

The following volumes apply to a 20µl DNA ligation.

add the following components to PCR tube: 
*'''9µl''' DNA plasmids
*'''1µl''' biobricks
*'''10ul''' Solution I (T4 ligase and buffer)
*'''MIX''' the reaction by stiring 
*Incubate the reaction at 16*C for 2hrs or overnight to ensure complete ligation.