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RACE problems - 5-terminal amplication (Nov/27/2006 )

My wife was dedicated in cloning of the whole genome of virus. the 5-terminal can not be acquired, because it is full of secondary structure. through her hard work, the secondary structure has been broken.
PROBLEM:she use terminal transferase which add poly(T) to the 5-terminal, then poly(A) and a specific antisense is used to amplify the 5-terminal. it does not work.

the protocol is ok? or you have a better protocol to get the 5-terminal of a fragment.


You might consider the SMART technique