Ligation

Materials:

• H₂O
• 1.5ml tubes
• Ligation buffer
• Ligase
• Insert
• Vector

Reaction:

Component	30μl reaction	Final concentration
H2O	up to 30μl
Buffer 10x	3μl	1x
DNA	4μl	up to 1μg
Enzyme/s	0.5μl

All the reagents are added following the order listed in the table above.
After the reaction is ready mix the content of the tube and spin it down.
The tubes are incubated for 1h at 37°C.

Reference: http://www.thermoscientificbio.com/uploadedFiles/Resources/fast-digestion-dna.pdf