To test the efficiency of the parallel cloning, plasmids from 28 individual colonies were isolated and the sequence of the inserts were determined by Sanger sequencing. Out of the 28 colonies, 25 contained inserts without errors; of those, 24 were unique sequences suggesting that the cloning efficiency was high and there was negligible sequence-bias in the plasmid pool. Furthermore, as described below, all 200 sequences were observed in the NGS of the entire pool.
