PNK Treatment of DNA Ends protocol

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Solutions/reagents:

• PNK stock
  
  (10, 000 U/ml)
• T4 ligase buffer
• substrate

Equipment:

• Incubator
• Reaction tubes

Steps:

1. Reaction Mix (10 µl)
   Use the following table as a checklist for preparing the reaction in reaction tube (1):
   
   

   	PNK stock	T4 ligase buffer	substrate
   Reaction Mix	1 µl	1 µl	8 µl

2. Reaction Conditions
   Incubate at 37°C for 30 mins.
   Incubate at 65°C for 20 mins.
   Store the tube on ice.
   

TOTAL TIME REQUIRED FOR THE COMPLETION OF THE PROTOCOL :~ 50 mins