SEQUENCING 500 KB OF YEAST DNA USING A GATC(R)-1500 IRECT BLOTTING ELECTROPHORESIS SYSTEM

We have developed efficient protocols Sor the construction of shotgun libraries and for high-throughput DNA sequencing at low cost using the optimized GATC(R) 1500 Direct Blotting Electrophoresis system with co lorimetric detection on nylon membranes. We can now sequence entire co smids without any primer walking at low coverage (8-fold to 9-fold). W e have demonstrated the efficiency of our protocols while sequencing 5 00 kb of yeast DNA with the GATC 1500 system. The methods described he re have allowed ur to achieve the highest data rate within the framewo rk of the European communities yeast sequencing program In 1993, only two people were involved in generating and submitting 156 274 bp of fi nal sequence data. Thereof 39 360 bp have been verified independently in other laboratories, and no sequencing error has been found.