SENSITIVE AND FAST MUTATION DETECTION BY SOLID-PHASE CHEMICAL CLEAVAGE

We have developed a solid phase chemical cleavage method (SpCCM) for s creening large DNA fragments for mutations. All reactions can be carri ed out in microtiterwells from the first amplification of the patient (or test) DNA through the search for mutations. The reaction time is s ignificantly reduced compared to the conventional chemical cleavage me thod (CCM), and even by using a uniformly labelled probe, the exact po sition and nature of the mutation can be revealed. The SpCCM is suitab le for automatization using a workstation to carry out the reactions a nd a fluorescent detection based DNA sequencing system to analyze the cleaved fragments. (C) 1996 Wiley-Liss, Inc.