| abstract |
An improved method of representational difference analysis (RDA) is described. Samples of tester and driver nucleic acids are compared by ligating a sample of the tester nucleic acids with a first adaptor, ligating a sample of the driver nucleic acids with a second adaptor and combining said ligated samples. Said combined sample is denatured and then annealed to allow double stranded nucleic acid molecules to reform. It is then digested with a restriction enzyme which preferentially digests hybrid tester:driver nucleic acid molecules at a restriction enzyme site generated by the interaction between the sequences of the first primer and the second primer attached to nucleic acids present in both tester and driver samples. Polymerase chain reaction (PCR) is used to amplify undigested tester sample nucleic acids. These steps may be repeated to generate further enriched samples. Kits, primer and adaptors for such methods are also claimed. |