Methods, systems, and arrays for biomolecular analysis
Disclosed herein are formulations, substrates, and arrays. Also disclosed herein are methods for manufacturing and using the formulations, substrates, and arrays. Also disclosed are methods for identifying peptide sequences useful for diagnosis and treatment of disorders, and methods for using the peptide sequences for diagnosis and treatment of disorders, e.g., celiac disorder. In certain embodiments, substrates and arrays comprise a porous layer for synthesis and attachment of polymers or biomolecules.
1. A method of assuring uniformly high quality features that are affixed to a surface of a microarray at positionally-defined locations, comprising:
a) coating the microarray with a coupling formulation comprising the features;
b) soft-baking the coated microarray;
c) determining a thickness of the soft-baked microarray;
d) stripping off the coat if the thickness of the soft-backed microarray falls outside a first threshold range, and repeating steps (a) through (c);
e) exposing the soft-baked microarray to light under a photomask;
f) hard-baking the exposed microarray; and
g) stripping off the coat if the thickness of the hard-baked microarray falls outside a second threshold range, and repeating steps (a) through (g).
2. The method of claim 1 , wherein the photomask comprises a diffusion pattern and an overlay pattern.
3. The method of claim 2 , further comprising the steps of
h) determining a percent deviation of the diffusion pattern or the overlay pattern of the hard-baked microarray as compared to a standard test diffusion or overlay pattern; and
j) optionally stripping off the coat if the percent deviation of the diffusion pattern or the overlay pattern is greater than 5%;
and repeating steps (a) through (i).
4. The method of claim 1 , wherein the first threshold range is from 2370 nm to 2430 nm.
5. The method of claim 1 , wherein the second threshold range is from 790 nm to 810 nm.
6. The method of claim 1 , wherein the microarray is soft-baked at a temperature from 75° C. to 115° C.
7. The method of claim 6 , wherein the microarray is soft-baked at 85° C.
8. The method of claim 1 , wherein the microarray is soft-baked from between 60 seconds to 120 seconds.
9. The method of claim 8 , wherein the microarray is soft-baked for 90 seconds.
10. The method of claim 1 , wherein the exposing step comprises a light at 248 nm.
11. The method of claim 1 , wherein the microarray is hard-baked at a temperature from 75° C. to 115° C.
12. The method of claim 11 , wherein the microarray is hard-baked at 85° C.
13. The method of claim 1 , wherein the microarray is hard-baked from between 60 seconds to 120 seconds.
14. The method of claim 13 , wherein the microarray is hard-baked for 90 seconds.
15. The method of claim 1 , wherein the coupling formulation comprises a photoactive compound, a polymer, and a coupling molecule.
16. The method of claim 15 , wherein the photoactive compound comprises a photobase.
17. The method of claim 15 , wherein the coupling molecule comprises an amino acid.
18. The method of claim 15 , wherein the coupling formulation further comprises a coupling reagent and a solvent.