In our lab we are developing a microtiter plate-based assay for the
detection of PCR amplified DNA fragments. One of the two primers is
labeled with biotine and is captured by a streptavidin coated
microtiter plate. After hybridization with a digoxigenin labeled probe
the label is detected using an enzyme immuno assay.
Our problem is that so far we have not been able to find a suitable
washing buffer that can be used in all washing steps throughout the
procedure. We tried buffers based on PBS and SSC but were not able to
detect any product. A washing buffer which is part of a commercial kit
gives exellent results but the composition is not revealed by the
Is there anyone who uses a similar assay and has a recipe of a
suitable wahing buffer?
Any help would be greatly appreciated.
H. Wilke (galaxy at pi.net)