AFFINITY TAG NUCLEIC ACID AND PROTEIN COMPOSITIONS AND PROCESSES FOR USING SAME
The present invention concerns compositions and processes that use affinity tags for isolating, and detecting or quantifying analytes, including nucleic acids, proteins and polypeptides. Compositions include nucleic acid compositions and protein compositions with affinity binding pairs, including metal binding peptides and immobilized metals, or peptide affinity groups.
1 . A composition of matter, comprising:
a chimeric molecule comprising a polynucleic acid portion and a peptide affinity tag portion covalently attached to the polynucleic acid portion; and
a nucleic acid hybridization and affinity tag binding buffer solution comprising a polyamine,
wherein the chimeric molecule is dissolved in the nucleic acid hybridization and affinity tag binding buffer.
2 . The composition of claim 1 , wherein the polyamine comprises putrescine.
3 . The composition of claim 1 , further comprising a water exclusion agent.
4 . The composition of claim 2 , further comprising a water exclusion agent.
5 . The composition of claim 1 , further comprising a polyethylene glycol (PEG).
6 . The composition of claim 2 , further comprising a polyethylene glycol (PEG).
7 . The composition of claim 1 , further comprising dextran sulfate.
8 . The composition of claim 2 , further comprising dextran sulfate.
9 . The composition of claim 1 , further comprising ethanol.
10 . The composition of claim 2 , further comprising ethanol.
11 . The composition of claim 1 , wherein the peptide affinity tag portion comprises polyhistidine.
12 . The composition of claim 2 , wherein the peptide affinity tag portion comprises polyhistidine.
13 . The composition of claim 1 , wherein the polynucleic acid portion comprises DNA.
14 . The composition of claim 2 , wherein the polynucleic acid portion comprises DNA.
15 . The composition of claim 1 , wherein the polynucleic acid portion is a DNA oligonucleotide.
16 . The composition of claim 2 , wherein the polynucleic acid portion is a DNA oligonucleotide.
17 . The composition of claim 1 , further comprising a first polynucleic acid molecule hybridized to the polynucleic acid portion of the chimeric molecule.
18 . The composition of claim 2 , further comprising a first polynucleic acid molecule hybridized to the polynucleic acid portion of the chimeric molecule.
19 . The composition of claim 1 , further comprising particles having a surface that specifically binds the affinity tag portion of the chimeric molecule.
20 . The composition of claim 2 , further comprising particles having a surface that specifically binds the affinity tag portion of the chimeric molecule.
21 . The composition of claim 19 , wherein the particles are magnetic or paramagnetic.
22 . The composition of claim 20 , wherein the particles are magnetic or paramagnetic.
23 . The composition of claim 1 , further comprising a test sample comprising a plurality of polynucleic acid molecules.
24 . The composition of claim 23 , wherein the test sample comprises a polynucleic acid molecule comprising a sequence complementary to the polynucleic acid portion of the chimeric acid molecule.
25 . The composition of claim 2 , further comprising a test sample comprising a plurality of polynucleic acid molecules.
26 . The composition of claim 25 , wherein the test sample comprises a polynucleic acid molecule comprising a sequence complementary to the polynucleic acid portion of the chimeric molecule.
27 . The composition of claim 1 , wherein the chimeric molecule is NAcHis-His-His-His-His-His-His-Cys-AGCAGGATTGCGGGTGCCAATGTGATCTTTAC.
28 . The composition of claim 2 , wherein the chimeric molecule is NAcHis-His-His-His-His-His-His-Cys-AGCAGGATTGCGGGTGCCAATGTGATCTTTAC.