Restriction digestion

Fast digest

Materials:

• MQ water
• 1.5ml tubes
• Fast digestion buffer
• Restriction enzymes
• DNA

Reaction:

Compound	20µl	Final concentration
MQ water	up to 20µl
10x fast digestion buffer	2µl	1x
restriction enzyme(s)	1µl	1U
DNA	4µl	up to 1µg

When the reaction mix is ready, the content is mixed and incubated at 37°C for 1h.

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

Normal digest

Materials:

• MQ water
• 1.5ml tubes
• Enzyme digestion buffer
• Restriction enzymes
• DNA

Reaction:

Compound	20µl	Final concentration
MQ water	up to 20µl
10x restriction digestion buffer	2µl	1x
restriction enzyme(s)	0.5µl	1U
DNA	2µl	0.2-1.5µg

When the reaction mix is ready, the content is mixed and incubated at 37°C for 2h.