Hi, everyone.
one of my project is to couple the amino-modified PCR products to the Carboxyl magnetic beads. Now the coupling experiment has been finished. for calculating the coupling efficiency, I want to break the amido bond and make the amino-modified PCR products separate from the carboxyl magnetic beads. the method has been conducted that the heat treat of the complex of the PCR product and magnetic beads under alkaline conditions. However the results show the amino bond has not been broken. are there better methods to break the amino bond?
Best wishes.
You may try acid hydrolysis which is another classical method for hydrolysis of the peptide bond, usually it uses 6M HCl at 110C in vacuo for 24 h.
Perhaps you can quantify the PCR products without separating them from the beads. You could use a fluorescent DNA dye, for example, and compare the fluorescence to a DNA standard curve (with unmodified beads added to account for any background fluorescence from them).
- Badges
- Science method