PCR free sample technical repeat #5
Libraries PCR_FREE_[1-19] (SRR12495846 - SRR12495864) are technical repeats and their reads were merged before analysis. 25 ng of purified target DNA samples (plasmid or PCR product) were premixed with 0.6 Âµl of the MuA UMI-transposome complex in 30 Âµ l of the tagmentation buffer. Tagmentation was carried out for 5 minutes at 30C, then stopped by adding 3 Âµl of 4.4% SDS solution. The tagmented DNA was subjected to the purification by modified size selection protocol using the Invitrogen Collibri DNA Library Cleanup Kit (Thermo Scientific), followed by PCR amplification with Invitrogen Collibri Library Amplification Master Mix (Thermo Scientific) using primers 5-AATGATACGGCGACCACCGAGATCTACAC[i5]ATGCGACACTCGTGAAACGCTTTCGCGTTT and 5-CAAGCAGAAGACGGCATACGAGAT[i7]CGAGGTCGAGTGCATGAAACGCTTTCGCGTTT, where [i5] and [i7] are library barcodes.