Oligonucleotide sequence formula for labeling olignucleotide probes and proteins for in-situ analysis
The present invention provides oligonucleotide probes and oligonucleotide probe collections and protein labeling for detecting or localizing a plurality nucleic acid target genes or antigens 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 provides collections or “cocktails” of oligonucleotide probes for detecting or localizing specific nucleic acid target genes within a cell or tissue sample. The cocktails are useful for detecting the following: the Kappa gene (SEQ ID NOS: 1-16 inclusive); the Lambda gene (SEQ ID NOS: 17 through 29 inclusive); the CMV (cytomegalovirus) gene (SEQ ID NOS: 30 through 50 inclusive); EBER (Epstein-Barr early RNA) gene (SEQ ID NOS: 51-54 inclusive); Alu (SEQ ID NOS: 55-56); PolyA (SEQ ID NO: 57); and the detection tail (SEQ ID NO: 58).
1. A method for preparing a synthetic oligonucleotide probe, comprising:
preparing an oligonucleotide molecule, wherein the oligonucleotide molecule comprises:
a first domain consisting of the sequence 5′-(CTATTTT) n CT-3′, wherein “n” is at least four and the first domain is detectably labeled through at least one cytosine in the sequence; 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,
thereby preparing the synthetic oligonucleotide probe.
2. The method of claim 1 , wherein the first domain is located 5′ of the second domain or the first domain is located 3′ of the second domain.
3. The method of claim 1 , wherein the first domain is detectably labeled with a reporter molecule or a hapten.
4. The method of claim 3 , wherein the hapten is fluorescein linked to the N4 nitrogen of cytosine through an N 4 -[2,2,-oxy-bis-(ethylamine)]-2′-deoxycytidine-5′-nucleotide linker.
5. The method of claim 3 , wherein the reporter molecule is a fluorophore.
6. A method for preparing a synthetic oligonucleotide probe, comprising:
preparing an oligonucleotide molecule, wherein the oligonucleotide molecule comprises:
a first domain consisting of the sequence 5′-(CTATTTT) n CT-3′, wherein “n” is at least four and the first domain is detectably labeled through at least one cytosine in the sequence; 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, and wherein the second domain comprises a sequence selected from the group consisting of SEQ ID NOs: 59-114.