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Supplementary data

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posted on 17.10.2019 by Dirk Höper, Leonie F. Forth

Table S1. Oligonucleotide sequences used in this study

Figure S1. Library yield depending on the number and position of phosphorothioate bonds in the

adapter oligonucleotides. The used adapters contained different numbers of phosphorothioate

bonds at different positions within the oligonucleotide. Oligonucleotide designations: BC, barcode number of the Ion Xpress index sequence; pt, adapter oligonucleotide containing given number of

phosphorothioate bonds; bc, phosphorothioate bonds located upstream of the barcode within the

key sequence. For further details and the oligonucleotide sequences, see Table 1 in the main

document. All libraries were prepared of the same batch of fragmented E. coli DNA in two independent experiments; vertical lines indicate the standard deviation calculated from three dilutions used in qPCR quantification. The horizontal line indicates the mean concentration of all libraries shown in the plot, the dashed lines indicate the mean ± 1 standard deviation

Figure S2.

Melting curve analysis of libraries prepared using standard-and Y-adapters. Melting curves were recorded after qPCR quantification for 0ng, 2 ng, 10 ng, and 50 ng DNA input. In comparison with the reference curves, the presence of adapter dimers is clearly visible. Reference was assembled of single peaks for non-template control (primer dimer; yellow), library without input (adapter dimer; red) and library quantification standard (153 bp; black).

Funding

This research was funded by the German Federal Ministry of Education and Research within project “DetektiVir” [grant number 13N13783]. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed. No writing assistance was utilized in the production of this manuscript.

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