Oligonucleotide sequence formula for labeling oligonucleotide probes and proteins for in-situ analysis
The present invention provides oligonucleotide probes and oligonucleotide probe collections for detecting or localizing a plurality nucleic acid target genes within a cell or tissue sample. Specifically, the invention provides collections of oligonucleotide probes for use in in situ hybridization analyses in which each probe has a label-domain with the sequence formulas of (CTATTTT) n CT, (AAAATAG) n or (TTTTATC) n or (GATAAAA) n in which all cases “n” would equal 1 or greater. The present invention also provides collections or “cocktails” of oligonucleotide probes for detecting or localizing specific nucleic acid target genes within a cell or tissue sample.
1. A synthetic oligonucleotide probe having a 5′ end and a 3′ end and comprising:
a first domain consisting of the sequence 5′-(AAAATAG) n -3′ wherein “n” is at least four, wherein the first domain does not hybridize with human nucleic acids under standard stringency conditions, wherein the first domain is detectably labeled, and wherein the 5′ end of the probe consists of the first domain; and
a second domain comprising a target sequence capable of hybridizing to a DNA or RNA sequence, wherein the second domain is not detectably labeled, the beginning of the second domain starting at the 3′ end of the first domain and the end of the second domain being the 3′ end of the probe, wherein the first domain and second domain comprise a single polynucleotide molecule.
2. The synthetic oligonucleotide probe of claim 1 , wherein the label is a fluorophore or a hapten.
3. The synthetic oligonucleotide probe of claim 2 , wherein the fluorophore is present at a density of greater than 7 mole percent of the first domain.
4. A synthetic oligonucleotide probe having a 5′ end and a 3′ end and comprising:
a first domain consisting of the sequence 5′-(AAAATAG) n -3′ wherein “n” is at least four, wherein the first domain does not hybridize with human nucleic acids under standard stringency conditions, wherein the first domain is detectably labeled, and wherein the 3′ end of the probe consists of the first domain; and
a second domain comprising a target sequence capable of hybridizing to a DNA or RNA sequence, wherein the second domain is not detectably labeled, the beginning of the second domain starting at the 5′ end of the probe and the end of the second domain being at the 5′ end of the first domain, wherein the first domain and second domain comprise a single polynucleotide molecule.
5. The synthetic oligonucleotide probe of claim 4 , wherein the label is a fluorophore or a hapten.
6. The synthetic oligonucleotide probe of claim 5 , wherein the fluorophore is present at a density of greater than 7 mole percent of the first domain.