3. 3. Multiplex LASSO cloning of the E. coli ORFeome
We next evaluated the ability of the new LASSO probes to capture a library of kilobase-sized ORFs from E. coli genomic DNA using the same capture parameters described by Tosi L. and coworkers (2017). According to the workflow in Fig 4a , LASSO probes were assembled as described above and hybridized with total genomic DNA of E.coli K12, targeting the 3078 ORFs in a single reaction volume. Circles containing ORFs were PCR amplified using primers that hybridize to the conserved adapter region on each LASSO probe.