Specific Oligodeoxynucleotide Probes Obtained through RNA Sequencing

Karl Köhrer, Toni M. Kutchan, Horst Domdey

Research output: Contribution to journalArticlepeer-review

11 Scopus citations


By combining several established techniques we developed a method to test the specificity of mixed oligodeoxynucleotide hybridization probes and to provide the information for the design of long nondegenerate, and therefore more specific probes. Mixed oligodeoxynucleotide probes derived from known peptide sequences are first used to initiate primer extension reactions with poly(A)+RNA as template in the presence of three dNTPs and one ddNTP to generate cDNA transcripts of defined lengths. Comparing the lengths of the cDNA transcripts with the possible nucleic acid sequence coding for the known oligopeptide indicates whether the oligodeoxynucleotide mix hybridizes predominantly to the RNA of interest. In a second step, the oligodeoxynucleotide mix with the highest specificity is used for indirect RNA sequence analysis. This confirms the specificity of the probe and provides information to design a long, highly specific oligodeoxynucleotide probe for the gene of interest. This simple two-step-procedure helps to circumvent the time-consuming procedures of subcloning and sequencing of cross-hybridizing fragments.

Original languageEnglish
Pages (from-to)143-147
Number of pages5
Issue number2
StatePublished - Mar 1989


Dive into the research topics of 'Specific Oligodeoxynucleotide Probes Obtained through RNA Sequencing'. Together they form a unique fingerprint.

Cite this