"Digestion and location of restriction sites"
Proposed activity:
Digestion of phage lambda DNA by fast-acting XhoI, EcoRI and HindIII enzyme (5 minutes) (made by students)
8-well agarose gel electrophoresis of phage lambda and lambda phage DNA digested with XhoI, EcoRI and HindIII
Interpretations of the results using the instructions to find the size of DNA bands obtained and locate the restriction sites
Composition:
To make 4 gels of 25 ml

• 1.6 g agarose
  
• 100 ml of TAE 10 buffer x pH =8.3 to dilute 10 times to get
  1,000 ml of buffer
  
• 106 μl of lambda phage DNA
  
• 8μl of XhoI restriction enzyme with rapid digestion
  
• 8μl of EcoRI restriction enzyme with rapid digestion
  
• 8μl of HindIII restriction digest enzyme
  
• 100 μl of fast-acting restriction enzyme buffer
  
• 250 μl of deposit blue
  Technical and educational instructions available on our website.
  
  Necessary material:
  DNA electrophoresis tank (for immersed gel)
  70-110 V, 20my generator
  Water Bath or Microwave
• Gloves
  Micropipettes from 5 to 50μl and adapted cones
  Conservation: 3 months
  DNA, restriction enzymes and digestion buffer: in the freezer
  Colour (protected from light), TAE buffer, sterile water: in the refrigerator