• Thaw all reagents on ice.
  • Assemble reaction mix into 50 µL volume in a microfuge tube.
  • Add reagents in following order: water, buffer, BSA, DNA template, restriction enzyme.
  • Gently mix by tapping tube. Briefly centrifuge to settle tube contents.
  • Prepare negative control reaction without template DNA.
  • Prepare positive control reaction with template of known cutting site corresponding to the restriction enzyme of choice.
  • Typical Incubation time and temperature is 37°C for 1 hour, though time and temperature will vary depending on restriction enzyme used.
  • Restriction enzymes are typically inactivated by incubation at high temperature. Incubation time and temperature is 65°C for 20 min, though time and temperature will vary depending on restriction enzyme used.
  • Analyze the results of your PCR reaction via gel electrophoresis.
Component
Final Concentration/amount
water
to 50 µL
buffer
1X
BSA
0.05 units/µL
DNA template
1 µg
Restriction enzyme
5-10 U per µg of DNA template (should not exceed 10% of reaction volume)

Restriction Digestion Design Tools

REQUEST A QUOTE

Do you like the current new website?

Hate

Dislike

Neutral

Like

Love

*