Patent ID: 7229798

Claim:
A process for parallel, transposon-mediated sequencing of DNA, the process comprising: i) providing one or more target DNA sequences, each inserted into a vector at an insertion site; ii) amplifying each target DNA-containing vector and pooling them, wherein each target DNA sequence is represented in the pool in an substantially equal amount per kb; iii) exposing the pool of target DNA-containing vectors to a selectable transposon, wherein the selectable transposon integrates into the target DNA-containing vectors at substantially random sites to form a pool of target DNA- and transposon-containing vectors; iv) transforming cells with the pool of target DNA- and transposon-containing vectors and isolating and growing a representative number of individual transformants into cultures under selection conditions; v) performing a polymerase chain reaction on DNA from each culture, wherein the polymerase chain reaction uses primers complementary to the 3′ ends of the vector sequence at the insertion site and has an extension time during each reaction cycle of the polymerase chain reaction sufficient to efficiently produce a DNA with the size of a full-length of the vector sequence but too short to efficiently produce a DNA with the size of a full-length of the vector sequence plus the transposon sequence; vi) measuring the amount of DNA produced in each polymerase chain reaction; vii) selecting cultures whose polymerase chain reactions produced substantial amounts of DNA above a cutoff value; and viii) sequencing the transposon flanking regions of the target DNA-containing vector from the selected cultures.