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.