PROCESSING OF CDNA AND GENOMIC KILOBASE-SIZE CLONES FOR MASSIVE SCREENING, MAPPING AND SEQUENCING BY HYBRIDIZATION

Efficient procedures for managing a large number of M13 or plasmid clo nes have been developed. In addition to picking, clones are directly a rrayed in multiwell plates by dispensing diluted transfor mation mixtu res. Metal pin arrays are used for fast inoculations of preparative pl ates filled by medium or by PCR mixture. Growth of M13 clones in multi well plates is optimized to obtain a consistently high yield, and a PC R protocol is defined for reliable amplification of several thousand M 13 ol plasmid inserts per day in BioOvens. Over 80000 cDNA inserts hav e been amplified. The phages or amplified inserts are spotted on nylon filters using an array of pins having a flat bottom, 0.3 mm in diamet er The procedures are suitable for an automated processing of hundreds of thousands of short clones from representative cDNA and genomic lib raries. Hybridization of arrayed clones with oligonucleotide and compl ex probes can simplify the search for new genes and accelerate large-s cale sequencing.