OLIGONUCLEOTIDE LIBRARIES FOR HIGH THROUGHPUT DNA SEQUENCING

A PROPRIETARY MULTIPLE-DNA SYNTHESIS INSTRUMENT IS EMPLOYED IN PHASE I OF THIS PROJECT TO SYNTHESIZE THE LIBRARY OF 3,314 NONAMER OLIGONUCLEOTIDES AS SPECIFIED FOR USE IN HIGH THROUGHPUT GENOME SEQUENCING BY A CONTINUOUS "PRIMER WALKING" APPROACH. THE OLIGONUCLEOTIDES ARE SYNTHESIZED AT A RATE OF 100 PER DAY, AFFINITY-PURIFIED BY RPL REVERSE-PHASE CARTRIDGE CHROMATOGRAPHY, QUANTIFIED AND ALIQUOTED INTO A "STOCK LIBRARY" (APPROXIMATELY 99% OF THE MATERIAL) AND A "TEST LIBRARY" (APPROXIMATELY 1% OF EACH NONAMER, OR 1.5 MUG/TUBE, SUFFICIENT FOR 30 SEQUENCING REACTIONS PER PRIMER). THE TEST LIBRARY IS SHIPPED TO THE UPJOHN COMPANY FOR TESTING OF A PROPOSED HIGH THROUGHPUT SEQUENCING STRATEGY.