METHOD FOR GENOME SEQUENCING

摘要

The present invention relates to a high throughput method for the identification and detection of molecular markers wherein restriction fragments are generated and suitable adaptors comprising (sample-specific) identifiers are ligated. The adapter-ligated restriction fragments may be selectively amplified with adaptor compatible primers carrying selective nucleotides at their 3′ end. The amplified adapter-ligated restriction fragments are, at least partly, sequenced using high throughput sequencing methods and the sequence parts of the restriction fragments together with the sample-specific identifiers serve as molecular markers.

主权项

1. A method for genome sequencing, comprising:
(a) digesting artificial chromosome clones in a plurality of pools with one or more restriction endonucleases to provide for a set of restriction fragments for each pool, and wherein each clone is present in more than one pool; (b) ligating adaptors to one or both sides of the restriction fragments, wherein at least one adaptor contains a pool-specific identifier or a degenerate identifier section, respectively, to provide adaptor-ligated restriction fragments; (c) sequencing at least the pool-specific identifier and part of the restriction fragment; (d) assigning the restriction fragment sequences of step (c) to the corresponding clonal sources using the pool-specific identifiers; (e) generating a physical map by ordering the restriction fragment sequences; and (f) applying the physical map for assembling a genome sequence.