Quantizing immobilization - any shortcuts?

REE700A at MAINE.MAINE.EDU REE700A at MAINE.MAINE.EDU
Fri Sep 4 12:44:51 EST 1992


   We are attempting to optimize the immobilization of the third
strand in a triple helix assay study (although that is immaterial
to the question at hand).  We are using aminopropyl trimethoxy
silane in a toluene solution and are trying to optimize this step.
  We are studying both solution and vapor phase deposition (wrt
concentration, time and technique) for optimum silane coverage on
lithium niobate, lithium tantalate and quartz.
  Obviously, we can use glutaraldehyde and an enzyme as a fairly
quick test for amino groups on the crystal surface.  OTOH, a
quicker test is desireable.  We have dozens (potentially hundreds)
of 2mm x 2mm x .5mm chips which we want to quantize for available
amino groups.  Any quick, reasonable quantitative analysis would be
appreciated.

  In anticipation of the next steps, a similar quantitative test
for carboxyl groups is also desired.
  Also, a good spacer polymer between the probe and its pendant
amino (artifically included) anchor spot is desired.  Are dummy
(noncomplementary) bases good for this or do they interfere with
affinity? (I am not the biochem person in this project - just the one
with net access).  The probe is complementary to an internal part of
the target DNA and I am concerned about "tight" immobilization
interfering with probe activity.

   Although I say Quantitative, I mean relative...

   Please reply via e-mail to :   Jeff Andle
                                  Biode, Inc. & University of Maine
                                  "Because 2 checks R better than 1"

   REE700A at Maine.maine.edu        DoD #3005



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