High throughput direct end sequencing of BAC clones

Libraries constructed in bacterial artificial chromosome (BAC) vectors have become the choice for clone sets in high throughput genomic sequencing pro jects primarily because of their high stability. BAC libraries have been pr oposed as a source for minimally overlapping clones for sequencing large ge nomic regions, and the use of BAC end sequences (i.e. sequences adjoining t he insert sites) has been proposed as a primary means for selecting minimal ly overlapping crones for sequencing large genomic regions. For this strate gy to be effective, high throughput methods for BAC end sequencing of all t he clones in deep coverage BAC libraries needed to be developed. Here we de scribe a low cost, efficient, 96 well procedure for BAC end sequencing. The se methods allow us to generate BAC end sequences from human and Arabidopos is libraries with an average read length of >450 bases and with a single pa ss sequencing average accuracy of >98%. Application of BAC end sequences in genomic sequencing is discussed.