I have been trying to immobilize DNA by the bioconjugation of biotin and streptavidin, but I cannot get this work.
I added EDC and streptavidin to
COOH functionalized resin, reacted for 3 hours in room temperature, and then washed with PBS.
Then I tried to anneal biotin-labeled primer and FAM-labeled complementary sequence. (Heated it up to 95 degree then let it cool down to room temperature.
After I added SA-functionalized resin and FAM-Biotin strands together, the conjugation wouldn't work. The resins are not fluorescent after wash.
I also tried to use
PCR products which are labeled on both side.
Is there any problem with my assay? How long it takes for the conjugation of biotin and SA at very low concentration?
Thank you for any information.