Table 1 Comparison of standard versus improved solution hybrid selection methods
| Â | Manual standard SHS protocol | Automated improved SHS protocol | ||
|---|---|---|---|---|
Process step | Standard method | Drawbacks | Improved method | Advantages |
Shearing of genomic DNA | Covaris S2 | Single sample | Optimized Covaris E210 | Multi-sample, improved yield, tight size range |
Enzymatic cleanups | Individual spin columns | Low throughput, 50 to 60% recovery, manual | 'With-bead' SPRI | High throughput, 80 to 90% recovery, automated |
Solution hybrid selection capture | Manual, column- based | Labor intensive (6 samples/FTE/week) | Fully automated | Walkaway, high throughput (1,200 samples/4FTE/week) |
Final PCR enrichment | Denature, followed by PCR | Sample loss through transfers | Direct 'off-bead' PCR | Improved final yield |
In process quality control checkpoints | Agilent Bioanalyzer | Limited visibility until sequence results | Many | In process results: key predictors of sample, library and sequencing quality |