Optical reporter compositions
First Claim
Patent Images
1. A composition of the formula I:
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(cOpR)-[L-(NA)]m
Iwherein;
m is an integer greater than 1;
(cOpR) is a coated optical reporter particle comprising an upconverting phosphor (UCP) of the formula Na (YxYbyErz) F4;
(NA) is a nucleic acid oligomer of detectable sequence, said (NA) is a double stranded DNA molecule having a length of between 40 base pairs and 1000 base pairs; and
L is a linking group covalently bound to the coated optical reporter particle and to the nucleic acid oligomer, said linking group L comprises an alkylene moiety having a first end covalently bound to the coated optical reporter particle and a second end covalently bound to the nucleic acid oligomer.
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Abstract
This invention provides compositions that have a light emitting reporter linked to biomolecules, preferably, nucleotide oligomers. The light reporter particles are silylated and functionalized to produce a coated light reporter particle, prior to covalently linking the biomolecules to the light reporter particle. The light reporter particles of the invention can be excited by a light excitation source such as UV or IR light, and when the biomolecule is DNA, the attached DNA molecule(s) are detectable by amplification techniques such as PCR.
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Citations
15 Claims
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1. A composition of the formula I:
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(cOpR)-[L-(NA)]m
Iwherein; m is an integer greater than 1; (cOpR) is a coated optical reporter particle comprising an upconverting phosphor (UCP) of the formula Na (YxYbyErz) F4; (NA) is a nucleic acid oligomer of detectable sequence, said (NA) is a double stranded DNA molecule having a length of between 40 base pairs and 1000 base pairs; and L is a linking group covalently bound to the coated optical reporter particle and to the nucleic acid oligomer, said linking group L comprises an alkylene moiety having a first end covalently bound to the coated optical reporter particle and a second end covalently bound to the nucleic acid oligomer. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15)
y is from about 0.05 to about 0.35; and z is from about 0.1 to about 0.001.
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3. The composition of claim 1, wherein L is of the formula:
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-A-R1—
B—wherein; R1 is C2-8alkylene; -A- is a group covalently bonded to the surface of the coated optical reporter; and —
B—
is a group covalently bonded to the 3′
or 5′
end of the nucleic acid oligomer.
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4. The composition of claim 3, wherein -A- is —
- O—
.
- O—
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5. The composition of claim 3, wherein —
- R1—
is —
(CH2)n- and wherein n is from 2 to 8.
- R1—
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6. The composition of claim 3, wherein —
- B—
is;—
S—
;—
O—
;—
NRa—
;—
S—
(CH2)p—
;—
O—
(CH2)p—
;—
NRa—
(CH2)p—
;—
S—
(CH2)q—
C(O)—
NRa—
(CH2)p—
;—
O—
(CH2)q—
C(O)—
NRa—
(CH2)p—
;—
NRa—
(CH2)q—
C(O)—
NRa—
(CH2)p—
;—
S—
C(O)—
(CH2)r—
C(O)—
NRa—
(CH2)p—
;—
O—
C(O)—
(CH2)r—
C(O)—
NRa—
(CH2)p—
;
or—
NRa—
C(O)—
(CH2)r—
C(O)—
NRa—
(CH2)p—
;wherein; p is from 2 to 8; q is from 1 to 8; r is from 2 to 8; and each Ra is independently hydrogen or C1-6alkyl.
- B—
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7. The composition of claim 3, wherein —
- B—
is;—
S—
(CH2)q—
C(O)—
NRa—
(CH2)p or—
N Ra—
C(O)—
(CH2)r—
C(O)—
NRa—
(CH2)p—
;wherein p, q, r and Ra are as recited in claim 6.
- B—
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8. The composition of claim 7, wherein:
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p is from 2 to 6; q is from 1 to 3; and r is 2 or 3.
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9. The composition of claim 3, wherein —
- B—
is;—
S—
CH2—
C(O)—
NH—
(CH2)6—
;
or—
NH—
C(O)—
(CH2)3—
C(O)—
NH—
(CH2)6—
.
- B—
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10. The composition of claim 1, wherein the cOpR is coated with silica.
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11. The composition of claim 10, wherein the silica comprises at least one Si—
- O bond.
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12. The composition of claim 1, wherein the NA is double stranded DNA molecule identifiable by real time PCR without DNA sequencing.
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13. The composition of claim 1, wherein the NA is a DNA molecule having a length of between 80 base pairs and 500 base pairs.
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14. The composition of claim 1, wherein said nucleic acid oligomer is identifiable by a method comprising polymerase chain reaction amplification while said oligomer is covalently attached to said coated optical reporter particle.
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15. The composition of claim 14, wherein said polymerase chain reaction amplification produces one or more amplicons and the size of said one or more amplicons is determined by a method comprising capillary electrophoresis.
Specification