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.