One-round conventional PCR

The 50-μl PCR mixtures contained 1 ng of equimolar mixture of plasmid#1 and plasmid#2 as template, 1 × Phusion HF Buffer (Thermo Fisher Scientific), 200 μM dNTPs, 0.5 μM primers Libr-A₁-for/Libr-rev (primer sequences are provided in Table 3) and 2 U of Phusion High-Fidelity DNA Polymerase (Thermo Fisher Scientific). Amplification was performed under the following conditions: 95 °C for 1 min; 25 cycles of 95 °C for 10 s, 55 °C for 30 s, and 72 °C for 10 s; 72 °C for 5 min. Aliquots of 10 μl of the reactions were analyzed by agarose gel electrophoresis.

Oligonucleotide primers used in the study