1. A method of expanding the set of distinguishable codes for encoding particles in a bioassay of a sample, where analytes are distinguished using encoded particles displaying different ligands, where ligands which bind to different analytes can be distinguished through the codes, comprising:
forming groups, each group made up of different numbers of particles having the same code, where members of each group display up to four different ligands, and wherein the number of particles in each group displaying the same ligand has a unique decomposition into one or more summands, such that no partial sum of one or more summands can be obtained in any other way of combining summands, and no summand is itself the sum of two or more of the other summands;
reacting the sample with the particles;
determining for each group of particles, the number of particles generating a positive assay signal;
comparing for each group of particles, said number with the number of particles displaying each said different ligand; and
determining for each group of particles, which analytes which bind to the ligands attached to the particles in a group, are present in the sample.
2. The method of claim 1 used to analyze the polymorphic form of a nucleic acid analyte present in a genomic sample, wherein each group of encoded particles is functionalized with one of four oligonucleotide probes, each probe differing by having one of the nucleotides A, C, G or T in the 3\u2032 terminal position.
3. The method of claim 2 wherein a positive assay signal is generated by an elongation reaction.
4. A method of expanding the distinguishable set of codes for coding particles displaying different ligands, each ligand being a peptide or an antibody, where the particles are used in a bioassay of a sample, and where ligands which bind to different analytes can be distinguished through the codes, comprising:
forming groups of different numbers of particles having the same code, where members of each group display up to four different ligands, and wherein the number of particles in each group displaying the same ligand has a unique decomposition into one or more summands, such that no partial sum of one or more summands can be obtained in any other way of combining summands, and no summand is itself the sum of two or more of the other summands;
reacting the sample with the particles;
determining for each group of particles, the number of particles generating a positive assay signal;
comparing for each group of particles, said number with the number of particles displaying each said different ligand; and
determining for each group of particles, which analytes which bind to the ligands attached to the particles in a group, are present in the sample.
5. The method of claim 4 wherein the ligands are peptides used to detect specific epitopes of auto-antibodies in a sample.
6. The method of claim 4 wherein determining the binding between peptides and antibodies is by a secondary detection antibody which binds to the auto-antibodies bound to the peptides.
7. The method of claim 4 wherein the solid phase carriers are beads encoded by different colors, and different colors correspond to different groups of peptide ligands, each such group of peptides corresponding to epitopes of a specific autoantibody.
8. The method of claim 4 wherein two or more different types of peptides are attached to beads of the same color.
The claims below are in addition to those above.
All refrences to claim(s) which appear below refer to the numbering after this setence.
1. A method for preparing a 1,3-benzoxazine comprising:
(a) combining a phenol compound, a diamino compound, an aldehyde compound, and water, to form a reaction mixture; and
(b) heating the reaction mixture for a time sufficient to form the benzoxazine.
2. The method of claim 1, wherein the reaction mixture further comprises an organic solvent.
3. The method of claim 2, wherein the amount of organic solvent is from about 1% to about 10%, by weight of the reaction mixture.
4. The method of claim 2, wherein the amount of organic solvent is from about 1% to about 5%, by weight of the reaction mixture.
5. The method of claim 2, wherein the amount of organic solvent is from about 1% to about 3%, by weight of the reaction mixture.
6. The method of claim 2, wherein the organic solvent is an alkyl acetate.
7. The method of claim 2, wherein the organic solvent is ethyl acetate.
8. A method for preparing a 1,3-benzoxazine comprising:
(a) combining a phenol compound, a diamino compound, an aldehyde compound, and water to form a reaction mixture; and
(b) heating the reaction mixture for a time sufficient to form the benzoxazine, wherein the reaction mixture further comprises a catalyst.
9. The method of claim 8, wherein the catalyst is a base.
10. The method of claim 9, wherein the base is sodium hydroxide, potassium hydroxide, sodium carbonate, potassium carbonate, or a mixture thereof.
11. The method of claim 1, further comprising the step of pouring the reaction mixture onto water after step (b).
12. The method of claim 1, further comprising the step of washing the reaction mixture of step (b) with aqueous base.
13. The method of claim 1, further comprising the step of washing the reaction mixture of step (b) with water.
14. A method for preparing a 1,3-benzoxazine comprising:
(a) combining a phenol compound, a diamino compound, an aldehyde compound, and water to form a reaction mixture; and
(b) heating the reaction mixture for a time sufficient to form the benzoxazine, wherein the reaction mixture is heated to a temperature of 100\xb0 C. or less.
15. The method of claim 1, wherein the reaction mixture is heated to a temperature of 90\xb0 C.\xb15\xb0 C.
16. The method of claim 1, wherein the time sufficient to form the benzoxazine is five hours or less.
17. The method of claim 1, wherein the phenol compound is phenol, cresol, 2-bromo-4-methylphenol, 2-allyphenol, 1,4-aminophenol, phenolphthalein, biphenol, 4-4\u2032-methylene-di-phenol, 4-4\u2032-dihydroxybenzophenone, bisphenol-A, 1,8-dihydroxyanthraquinone, 1,6-dihydroxnaphthalene, 2,2\u2032-dihydroxyazobenzene, resorcinol, fluorene bisphenol, or 1,3,5-trihydroxy benzene.
18. The method of claim 1, wherein the aldehyde compound is formaldehyde, paraformaldehyde, or polyoxy.
19. A 1,3-benzoxazine produced by the method of claim 1.
20. The method of claim 1, wherein
the benzoxazine may be embraced by one or more of the following structures:
wherein o is 1-4, X is selected from direct bond (when o is 2), alkyl (when o is 1), alkylene (when o is 2-4), carbonyl (when o is 2), thiol (when o is 1), thioether (when o is 2), sulfoxide (when o is 2), and sulfone (when o is 2), R1 is selected from hydrogen, alkyl, alkenyl and aryl, and R4 is selected from hydrogen, halogen, alkyl and alkenyl; or
wherein p is 2, Y is selected from biphenyl (when p is 2), diphenyl methane (when p is 2), diphenyl isopropane (when p is 2), diphenyl sulfide (when p is 2), diphenyl sulfoxide (when p is 2), diphenyl sulfone (when p is 2), and diphenyl ketone (when p is 2), and R4 is selected from hydrogen, halogen, alkyl and alkenyl.