AUTOMATED 4-COLOR DNA-SEQUENCING USING PRIMERS ASSEMBLED BY HEXAMER LIGATION

A procedure based on the assembly of sequencing primers by hexamer lig ation and then using them in automated DNA sequencing is described. Th is method is based on a four-color fluorescent terminator chemistry. S equencing ladders were analyzed using an ABI 373 DNA sequencer (Applie d Biosystems, Foster City, CA, USA). The best results were obtained fo r primers assembled by ligation of four to ten hexamers. The accuracy of the method was estimated to be 99.5% up to 400 nt of the read seque nce, and somewhat lower at 400-600 nt.