Ligation

Materials:

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

Reaction:

Component	20μl reaction	Final concentration
H2O	up to 20μl
10x Ligation buffer	2μl	1x
Vector	1μl	20-100μg
Insert	1μl	1:1 molar ratio
T4 DNA Ligase 5U/μl	0.2μl	1U/μl

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 at room temperature (~22°C) for 1h.

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