Sequence scanning: A method for rapid sequence acquisition from large-fragment DNA clones

摘要

A strategy of "sequence scanning" is pro- posed for rapid acquisition of sequence from clones such as bacteriophage P1 clones, cosmids, or yeast artificial chromo- somes. The approach makes use of a special vector, called LambdaScan, that reliably yields subclones with inserts in the size range 8-12 kb. A number of subclones, typically 96 or 192, are chosen at random, and the ends of the inserts are sequenced using vector-specific primers. Then long-range spectrum PCR is used to order and orient the clones.