Self-assembling high density ordered patterned biomolecule array and method for making and using the same
A method for fabricating a universal substrate for attaching biomolecules, including sequencing features and the resulting substrate. A method of direct detection of analytes utilizes a Complementary Metal Oxide Semiconductor (CMOS) sensor with the substrate.
1. A patterned substrate comprising a substrate comprising a plurality of first regions and a second region wherein:
the plurality of first regions have a density on the substrate between 5×10 6 and 25×10 6 /mm 2 and an associated biomolecule specifically located at each of said plurality of first regions and a complementary metal-oxide semiconductor (CMOS) sensor specifically located at each of said plurality of first regions.
2. The substrate of claim 1 wherein the associated biomolecule is a polynucleotide.
3. The substrate of claim 2 wherein the polynucleotide is clonally amplified.
4. The substrate of claim 1 wherein the associated biomolecule is a polypeptide.
5. The substrate of claim 1 further comprising a polymer attached to the second region.
6. The substrate of claim 3 wherein the clonally amplified polynucleotides attached to each of the plurality of first regions are different.
7. The substrate of claim 1 wherein each of the plurality of first regions are coupled to a CMOS pixel.
8. The substrate of claim 1 wherein the density is between 6×10 6 and 10×10 6 sequencing features/mm 2 .
9. A method for building a patterned substrate comprising:
providing a substrate comprising a plurality of complementary metal oxide semiconductor (CMOS) sensors at a density of between 5×10 6 and 25×10 6 /mm 2 ;
etching a well onto the substrate at each CMOS sensor;
coating the plurality of wells with a first coating; and
associating a biomolecule to each of the plurality of wells.
10. The method of claim 9 wherein the biomolecule comprises a polynucleotide.
11. The method of claim 9 wherein the biomolecule comprises a polypeptide.
12. The method of claim 10 wherein associating a polynucleotide to the plurality of wells to comprises:
attaching a universal primer to each of the plurality of wells;
hybridizing a circular DNA template onto each universal primer; and
performing solid-phase rolling circle amplification.
13. The method of claim 10 wherein associating a polynucleotide to the plurality of wells comprises:
attaching universal forward and reverse primers to each of the plurality of wells;
hybridizing DNA template onto one of the forward or reverse primer; and
performing solid-phase bridge amplification.
14. The method of claim 10 wherein associating a polynucleotide to the plurality of wells comprises:
attaching universal primer to each of the plurality of wells; and
hybridizing the polynucleotide comprising a DNA template to the universal primer.
15. The method of claim 10 wherein associating a polynucleotide to the plurality of wells comprises clonally amplifying the polynucleotide.
16. The method of claim 10 wherein the polynucleotide is a single DNA template.
17. The method of claim 10 wherein associating a polynucleotide to the plurality of wells comprises a self-assembling process.
18. The method of claim 9 wherein the density is between 6×10 6 and 10×10 6 sequencing features/mm 2 .