Team:Groningen/protocols/Digestion

Restriction digestion

Fast digest

Materials:

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

Reaction:

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

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

Normal digest

Materials:

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

Reaction:

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

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

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