Nonradiocative Northern Method Recomendation

Kajetan H. Groicher Kajetan-Groicher at ouhsc.edu
Tue Nov 2 16:47:04 EST 1999


I have used both the Digoxygenin system from Boehringer, and the
Alk-Phos direct system from Amersham (No assoc etc...).  Each kit has
it's advantages and disadvantages.  Namely: the Dig-kit has a few extra
steps in detection (essentially a Western step) but has more flexibility
in generating probes (such as random priming, PCR, klenow etc.)  The
Amersham kit is nice b/c the probe can be labeled in a matter of a half
an hour, and the signal generation/detection is less complex than the
Dig system.  The main disadvantage to the Amersham system (in my
opinion) is the lack of flexibility in post hyb stringency washes.  This
system utilizes an enzyme that is only stable to ~55oC, so high temp
washes are out of the question, although you do have great latitude in
adjusting alt concentrations to make up for it.

In my hands both kits have worked very well, the quirks associated with
each system of minor consequence.  Information and protocols for these
kits are available on line from the supplier.

Best of luck.

Kajetan

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