• Created by Unknown User (shinjini@mit.edu), last modified by Unknown User (krbrink@mit.edu) on Jul 28, 2015 13:06

Digestion Protocol

  • 20 ul total
  • 500-1000 ng DNA
  • 1 ul enzyme
  • 2 ul enzyme buffer
  • fill the rest with water
  • Pipette up and down thoroughly 
  • 37 degrees for 1-3 hours
  • 4-5 ul loading dye
  • gel + ladder!

From NEB:

One unit is defined as the amount of enzyme required to digest 1 µg of λ DNA in 1 hour at 37°C in a total reaction volume of 50 µl


So check calculate how much DNA you have and use the right amount of enzyme. Or more.

An excel document is attached to aid in making calculations for multiple parallel digests easier. Use this document and attach it to your experiment for science and stuff.

 

Lab enzyme chart (7/28/2015):

  • No labels