JROSENBAUM.CURRY@BIONET-20.BIO.NET (Alice Curry) (04/27/89)
Our lab is studying mutant cells that have been rescued by transformation with a wild type gene. The gene is one that undergoes transcriptional induction, and we want to study the induction of the transformed wild type copies. However, the mutant gene, which may still be intact in the transformants, undergoes normal induction. So mutant cells have plenty of normal-sized message, which will show up on the Northern and mask the message made by any transformed copies of the gene. To get around this problem, we want to use oligonucleotides as probes. We want to make two oligos, 17-mers that have the mutant nucleotide in the middle. (i.e., one oligo matches the mutant sequence at that point, one matches the wild type.) Because we've never used oligos as probes, and haven't yet found any information about detecting single nucleotide differences in RNAs (the papers we found so far discuss DNA:DNA conditions), we would appreciate any references and advice that people n give us regarding this type of experiment. Thanks. --Alice Curry, Yale U., Biology Dept. -------