Methods, kits and compositions pertaining to combination oligomers and libraries for their preparation
First Claim
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1. A kit comprising:
- (a) two or more independently detectable combination oligomers wherein,(i) each of said independently detectable combination oligomers comprises a first oligomer block and a second oligomer block that are each independently a peptide nucleic acid, a PNA chimera or PNA combination oligomer and are linked covalently to each other by a linker that is at least three atoms in length that is not part of a nucleobase containing backbone subunit of the polymer,(ii) in each independently detectable combination oligomer, the first and second oligomer blocks taken together encode a probing nucleobase sequence that is designed to sequence specifically hybridize juxtaposed to a target sequence of contiguous nucleobases, such that there is no gap or gap base, to thereby form a double stranded target sequence/combination oligomer complex, and the probing nucleobase sequence in each independently detectable combination oligomer differs from the probing nucleobase sequences of the other independently detectable combination oligomer(s) by at least one nucleobase, and(iii) each independently detectable combination oligomer contains at least one independently detectable label;
(b) optionally one or more oligonucleotides;
(c) optionally one or more buffers;
(d) optionally one or more nucleotide triphosphates;
(e) optionally a nucleic acid amplification master mix; and
(f) optionally one or more polymerase enzymes.
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Abstract
This invention pertains to the field of combination oligomers, including the block synthesis of combination oligomers in the absence of a template, as well as related methods, kits, libraries and other compositions.
44 Citations
32 Claims
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1. A kit comprising:
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(a) two or more independently detectable combination oligomers wherein, (i) each of said independently detectable combination oligomers comprises a first oligomer block and a second oligomer block that are each independently a peptide nucleic acid, a PNA chimera or PNA combination oligomer and are linked covalently to each other by a linker that is at least three atoms in length that is not part of a nucleobase containing backbone subunit of the polymer, (ii) in each independently detectable combination oligomer, the first and second oligomer blocks taken together encode a probing nucleobase sequence that is designed to sequence specifically hybridize juxtaposed to a target sequence of contiguous nucleobases, such that there is no gap or gap base, to thereby form a double stranded target sequence/combination oligomer complex, and the probing nucleobase sequence in each independently detectable combination oligomer differs from the probing nucleobase sequences of the other independently detectable combination oligomer(s) by at least one nucleobase, and (iii) each independently detectable combination oligomer contains at least one independently detectable label; (b) optionally one or more oligonucleotides; (c) optionally one or more buffers; (d) optionally one or more nucleotide triphosphates; (e) optionally a nucleic acid amplification master mix; and (f) optionally one or more polymerase enzymes. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16)
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17. A kit comprising:
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(a) two or more independently detectable combination oligomers wherein, (i) each of said independently detectable combination oligomers comprises a first oligomer block and a second oligomer block that are each independently a peptide nucleic acid, a PNA chimera or PNA combination oligomer and are linked covalently to each other by a linker that is at least three atoms in length and that is not part of a nucleobase containing backbone subunit of the polymer, (ii) in each independently detectable combination oligomer, the first and second oligomer blocks taken together encode a probing nucleobase sequence that is designed to sequence specifically hybridize to a target sequence of contiguous nucleobases to thereby form a double stranded target sequence/combination oligomer complex, and the probing nucleobase sequence in each independently detectable combination oligomer differs from the probing nucleobase sequences of the other independently detectable combination oligomer(s) by at least one nucleobase, and (iii) each independently detectable combination oligomer contains at least one independently detectable label; (b) optionally one or more oligonucleotides; (c) optionally one or more buffers; (d) optionally one or more nucleotide triphosphates; (e) optionally a nucleic acid amplification master mix; and (f) optionally one or more polymerase enzymes. - View Dependent Claims (18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32)
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Specification