Q re: PCR from agarose plugs

Mon Feb 3 20:47:14 EST 1997

Steven Sullivan wrote:
> WHat are the rules of thumb for re-eamplifying a band from a plug of
> agarose?  I have had only variable success doing this, often getting a
> smear or supernumerary bands instead of the target band alone. Is there a
> standard protocol somewhere?We've had good success using low melting temperature agarose (either an 
electrophoresis quality LMT-agarose for larger fragments or NuSieve-GTG 
for small fragments; melting to liquify; and diluting at least 5-fold 
into the second PCR reaction.

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