I am using TESPA (triethoxysilylpropyl succinic anhydride) to silanize glass surfaces for DNA functionalization.
I would like to add a blocking step to avoid non specific coupling of my target DNA.
Some blocking agent are described in litterature like ethanolamine, t-RNA, tween 20, BSA etc.
Did anybody try some of them ?
I would like to avoid optimization steps :)
Thanks in advance :)
How are you planning to carry out the DNA functionalization? Do you have amino-modified DNA?
Yes i do ! my capture sequence is modified with an amino group on its 5' end. I also use w signal sequence to check the hybridization. But this one seems to binds everywhere on the silanized surface .. this is why i am looking for a blocking agent :)
Do you mean that the signal sequence binds to the silica surface? It's possible under your reaction conditions that the exocyclic amines on your signal oligo can react with the succinic anhydrides on the silica surface. In that case, you can do a "backfill" after your initial functionalization with the capture sequence. I would recommend adding a small hydrophilic amine such as 2-[2-(2-aminoethoxy)ethoxy]ethanol to enure all succinic anhydride groups are inactivated. Good luck!
http://www.sigmaaldrich.com/catalog/product/aldrich/768901
Thanks a lot !! i'll keep this option, check and let you know :) thanks for your time again :)
- Badges
- Science topic
- Similar topics
- Chemistry
- Nanotechnology