Conditioned surfaces for in situ molecular array synthesis
Described herein are in situ synthesized arrays and methods of making them, wherein array signal sensitivity and robustness is enhanced by carrying out conditioning steps and/or generating linkers during synthesis. An array comprises a surface with a collection of features, wherein the features comprise molecules or polymers attached to the surface. In certain embodiments of the invention, carrying out conditioning steps during array synthesis can yield arrays with improved signal. In other embodiments, linkers are synthesized on the array surface prior to synthesis of functional molecules, wherein increasing linker length can correspond to an improvement in the signal generated by the array.
1. An array, comprising:
(a) a surface; and
(b) a plurality of molecules immobilized to the surface;
wherein the molecules comprise a peptide linker sequence having two ends, wherein one end of the peptide linker sequence is coupled to the surface and the other end is coupled to a functional sequence which binds a target molecule; and wherein the peptide linker sequence consists of GGSG, GGGSG, GGGGSG, GGGGGSG, or GGGGGGSG.
2. The array of claim 1 , wherein the peptide linker sequence consists of GGGGGSG.
3. The array of claim 1 , wherein the peptide linker sequence consists of GGGGGGSG.
4. An array, comprising:
(a) a surface; and
(b) a plurality of in situ synthesized molecules immobilized to the surface;
wherein the molecules comprise a peptide linker sequence having two ends, wherein one end of the peptide linker sequence is coupled to the surface and the other end is coupled to a functional sequence which binds a target molecule; and wherein the peptide linker sequence consists of GGSG, GGGSG, GGGGSG, GGGGGSG, or GGGGGGSG.
5. The array of claim 4 , wherein the peptide linker sequence consists of GGGGGSG.
6. The array of claim 4 , wherein the peptide linker sequence consists of GGGGGGSG.
7. A method of making an array, comprising:
(a) performing a chemical reaction upon the array that is capable of attaching an amino acid to a coupling site, wherein the amino acid forms part of a peptide linker sequence;
(b) performing an additional chemical reaction upon the array that is capable of adding an amino acid to the attached amino acid of step (a); and
(c) repeating step (b) at least once until the amino acids that comprise the desired peptide linker sequence have been added, wherein the desired peptide linker sequence consists of GGSG, GGGSG, GGGGSG, GGGGGSG, and GGGGGGSG;
(d) synthesizing in situ a functional sequence onto the free end of the peptide linker sequence, wherein the peptide linker sequence enhances binding between the functional sequence and a substrate.
8. The method of claim 7 , wherein the peptide linker sequence length consists of GGGGGSG.