Table 4. Library preparation workflow.
|
|
User hands-on time (min)
|
Incubation time (min)
|
Number of pipetting steps (for 16 samples)
|
||||
|---|---|---|---|---|---|---|---|
| Protocol step | Manual (1 sample) | Manual (16 samples)* | AMCC (16 samples) | Manual | AMCC | Manual | AMCC |
| Aliquoting samples/reagents | 5 | 20 | 16 | 81 | |||
| Concentrate sheared DNA | 30 | ||||||
| End repair | 10 | 20 | 30 | 30 | 4 (34)** | ||
| DNA purification | 30 | 30 | 30 | 80 | |||
| dA tailing | 10 | 20 | 30 | 30 | 4 (34)** | ||
| DNA purification | 30 | 30 | 30 | 80 | |||
| Adapter ligation | 10 | 20 | 15 | 15 | 5 (50)** | ||
| DNA purification | 30 | 30 | 30 | 80 | |||
| Sample transfer | 5 | 16 | |||||
| Total | 120 | 155 | 25 | 75 | 195 | 253 (358) | 97 |
Time and effort spent on library preparation for a conventional benchtop protocol compared with the on-chip workflow.
* Using a multichannel pipettor and microtiter plate for assembling enzymatic reactions
** Assumes that a master mix of enzyme plus buffer is assembled and then aliquoted with a multichannel pipettor. Numbers in parentheses indicate the number of pipetting steps if a standard pipettor is used.