detection of PCR products by hydridization in streptoavidin coated plates

Alejandro Palacios palaman at TERRABA.FING.UCR.AC.CR
Sat Aug 17 10:31:54 EST 1996

Hi neters,

I need help.

I am trying to make an in house PCR detection products system, we are 
interested in a plata hibridization based detction.  I have tryed to 
capture a biotinilated PCR product (only one strand labeled) in 
Sprettoavidin coated plates, after that denaturing with NaOH, 
hibridization with a DIG labeled probe and developing with antiDIG-AP 
with (pNPP).    
The problem is that everything in the tests, including negative controls

I suspect am not doing a good streptoavidin coating or there is too much 
affinity between pyloestherene and the probe. (0.1 microg/well in a 
IMULON 2 strip).

Is any specific way of coating and blocking plates for nucleic acid 
Is the concentration of streptoavidin too high?
Sugestion of other systems?
All comments welcome.

Alejandro Palacios         palaman at
International Center for Medical Research and Training (ICMRT)
Loussiana State University -  Costa Rica

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