Fig. 1

Flowchart for the All-in-One sequencing (AIO-seq) method. a The libraries were prepared using Tn5 transposase. b The process of mechanical fragmentation was used to prepare the libraries. c In the traditional protocol, the size selection, and quantification were processed using a one sample one tube method. d With the AIO-seq method, the library analyzed by the Agilent 2100 Bioanalyzer will give the fragment distribution pattern and the ratio of the target region (between the two blue lines) to the total library. e The concentration of the total library could be obtained by Qubit™ 4.0 Fluorometer. f The target region concentrations (TRC) were calculated within each library by multiplying the proportion of the target region from (d) and the total library concentration from (e). g Mixing the libraries in one tube according to the calculated TRC and their expected yields of the sequence data. h–i One size selection by Sage HT. j Quantification of the selected fragment by qPCR and sequencing