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What was the cloning method used for the C.elegans ORFs?
The C. elegans ORF clones started as a pool of amplimers from a cDNA library. For this reason there will be populational variation within this pool. Primer pairs were designed by computational methods, based on gene predictions from one of the first drafts of the worm genome annotation made public in 1999. Gene-specific primer pairs amplify all 'splice variants' between the two primers designed for any gene. Pools of ~50-1,000 transformants for each set of amplification products were partially sequenced to produce OSTs (ORF sequence tags). This confirmed gene identity and permitted identification of pools where at least one processed mRNA was represented.