Introduction
5’ overhangs can be end-labeled radioactively with T4 PolyNucleotide Kinase (PNK), or filled in using the Klenow fragment with labeled dNTPs:
-1ul T4 PNK
-1ul 32P ATP (3,000Ci/mmol, 5mCi/ml)
-2ul 10x T4 PNK buffer
-1 or 2ul DNA ladder (1ug)
Protocol
Add distilled water to a reaction volume of 20ul.
Incubate for 30 minutes at 37°C.
Run the samples for 50 to 60 minutes at 100V in TBE buffer in a 4-20% acrylamide gel (10cm x 10cm). A 20 minutes exposure gives very readable signals. The signal strength is about twice that signal when ADP is added to 100uM.
Choose your country
North America
Europe
Asia-Pacific
Session Expired
You have been idle for more than 20 minutes, for your security you have been logged out. Please sign back in to continue your session.
Institution Changed
Your profile has been mapped to an Institution, please sign back for your profile updates to be completed.
Sign in to your NEB account
To save your cart and view previous orders, sign in to your NEB account. Adding products to your cart without being signed in will result in a loss of your cart when you do sign in or leave the site.
