Table 1.
Library pooling of 14 rice samples using AIO-seq
| Group No. | Sample ID | Con. of lib (ng/μL)a | Ratio of 420–520 bp (%)b | Target region concentration (TRC, ng/μL) | Proportion of expected data yield in groupc | Mass of mixed target region (ng)c | Vol. for mixing (μL)d | Final data yield (Gb) |
|---|---|---|---|---|---|---|---|---|
| A | 1 | 21.00 | 11 | 2.31 | 1/7 | 20.0 | 8.66 | 6.38 |
| 2 | 25.60 | 13 | 3.33 | 1/7 | 20.0 | 6.01 | 6.02 | |
| 3 | 19.60 | 13 | 2.55 | 1/7 | 20.0 | 7.85 | 7.20 | |
| 4 | 20.40 | 12 | 2.45 | 1/7 | 20.0 | 8.17 | 6.97 | |
| 5 | 24.80 | 13 | 3.22 | 1/7 | 20.0 | 6.20 | 7.03 | |
| 6 | 22.20 | 12 | 2.66 | 1/7 | 20.0 | 7.51 | 6.50 | |
| 7 | 24.20 | 12 | 2.90 | 1/7 | 20.0 | 6.89 | 6.74 | |
| B | 1 | 20.20 | 10 | 2.02 | 1/7 | 20.0 | 9.90 | 5.79 |
| 2 | 25.80 | 13 | 3.35 | 1/7 | 20.0 | 5.96 | 5.88 | |
| 3 | 22.00 | 12 | 2.64 | 1/7 | 20.0 | 7.58 | 6.44 | |
| 4 | 24.60 | 12 | 2.95 | 1/7 | 20.0 | 6.78 | 5.95 | |
| 5 | 21.40 | 13 | 2.78 | 1/7 | 20.0 | 7.19 | 5.46 | |
| 6 | 24.60 | 13 | 3.20 | 1/7 | 20.0 | 6.25 | 5.96 | |
| 7 | 26.20 | 13 | 3.41 | 1/7 | 20.0 | 5.87 | 5.74 |
aCon. of lib means the initial concentration of library for each individual assayed by Qubit™ 4.0 Fluorometer
bAnalyzed by Agilent 2100 Bioanalyzer, indicates the ratio of the fragment between 420 and 520 bp, which will be size selected for sequencing
cFor each group, seven samples were processed together and each sample was expected to have an equal yield of data, thus the proportion of expected data yielded for each sample was 1/7, and we mixed equal amounts of the target regions (20.0 ng) for each
dVol. for mixing represents the volume of each library that needed to be pooled. Both the “Mass of mixed target region” and “Vol. for mixing” could be proportionately scaled up or down