Patent Document ID: 9150916
Application ID: 13532674
Patent Flag: 1

Claim One:
1. A method for determining genetic elements of an organism's genome, the method comprising: performing hyper-saturated transposon mutagenesis to generate an engineered transposon comprising a Tn5 O-end insertion primer having the sequence of SEQ ID NO: 3, and a Tn5 I-end primer having the sequence of SEQ ID NO. 2; said hyper-saturated transposon mutagenesis having a transposition density from about 8 to 100 bp; selecting for viable mutants in the engineered transposon under a given selection condition; pooling the selected viable mutants; isolating transposon junctions through a shared parallel PCR strategy utilizing a first primer that is transposon specific, at least one semi-arbitrary primer, and at least one adaptor sequence primer, wherein the at least one semi-arbitrary primer is selected from the group consisting of SEQ ID NO. 4, SEQ ID NO. 5, and SEQ ID NO. 6; sequencing the isolated transposon junctions in the ultra high-throughput adaptor based DNA sequencer; mapping transposon insertion sites onto the organism's genome; and identifying the genetic elements based on frequency, locations and insertion orientation of the mapped transposon insertion sites.