3.7 Sequencing
(See Notes 20 and 17)
1. Create Run using Local Run Manager Software (see Note 21).
From the home screen, select Edit Runs. Select “Create Run”
from the Local Run Manager dashboard and then select an
analysis module. Enter a run name, enter samples for the run,
and if applicable, import manifests. Save the run and close the
Local Run Manager dashboard window.
2. Remove the reagent cartridge from À25
C to À15
C storage.
3. Thaw cartridge in a room temperature water bath until thawed
(~60 min). Do not submerge the cartridge.
4. Gently tap on the bench to dislodge water from the base and
then dry the base.
5. Invert the cartridge five times to mix reagents.
6. Inspect positions 29, 30, 31, and 32 to make sure that reagents
are thawed.
7. Gently tap on the bench to reduce air bubbles.
8. Remove a new flow cell package from 2
C to 8
C storage.
9. Set the unwrapped flow cell package aside at room temperature
for 30 min. If the foil package is intact, the flow cell can remain
at room temperature up to 12 h. Avoid repeated cooling and
warming of the flow cell.
10. Remove the flow cell from the foil package.
11. Open the clear plastic clamshell package and remove the
flow cell.
12. Clean the glass surface of the flow cell with a lint-free alcohol
wipe. Dry the glass with a low-lint lab tissue.
13. Clean the foil seal covering reservoir #10 labeled “Load
Library Here” using a low-lint tissue.
14. Pierce the seal with a clean 1 mL pipette tip.
15. Load 1.3 mL of prepared 1.8 pM libraries (output from Subheading 3.6) into reservoir #10 labeled “Load Library Here”.
Avoid touching the foil seal as you dispense the libraries.
16. From the Home screen, select “Experiment.”
17. On the Select Assay screen, select “Sequence.” The Sequence
command opens the imaging compartment door, releases consumables from a previous run, and opens the series of run setup
screens. A short delay is normal.
18. Select one of the following run modes to determine where to
enter run information and how to analyze data (see Notes 22
and 23):
500
Shuyi Ma et al.
(See Notes 20 and 17)
1. Create Run using Local Run Manager Software (see Note 21).
From the home screen, select Edit Runs. Select “Create Run”
from the Local Run Manager dashboard and then select an
analysis module. Enter a run name, enter samples for the run,
and if applicable, import manifests. Save the run and close the
Local Run Manager dashboard window.
2. Remove the reagent cartridge from À25
C to À15
C storage.
3. Thaw cartridge in a room temperature water bath until thawed
(~60 min). Do not submerge the cartridge.
4. Gently tap on the bench to dislodge water from the base and
then dry the base.
5. Invert the cartridge five times to mix reagents.
6. Inspect positions 29, 30, 31, and 32 to make sure that reagents
are thawed.
7. Gently tap on the bench to reduce air bubbles.
8. Remove a new flow cell package from 2
C to 8
C storage.
9. Set the unwrapped flow cell package aside at room temperature
for 30 min. If the foil package is intact, the flow cell can remain
at room temperature up to 12 h. Avoid repeated cooling and
warming of the flow cell.
10. Remove the flow cell from the foil package.
11. Open the clear plastic clamshell package and remove the
flow cell.
12. Clean the glass surface of the flow cell with a lint-free alcohol
wipe. Dry the glass with a low-lint lab tissue.
13. Clean the foil seal covering reservoir #10 labeled “Load
Library Here” using a low-lint tissue.
14. Pierce the seal with a clean 1 mL pipette tip.
15. Load 1.3 mL of prepared 1.8 pM libraries (output from Subheading 3.6) into reservoir #10 labeled “Load Library Here”.
Avoid touching the foil seal as you dispense the libraries.
16. From the Home screen, select “Experiment.”
17. On the Select Assay screen, select “Sequence.” The Sequence
command opens the imaging compartment door, releases consumables from a previous run, and opens the series of run setup
screens. A short delay is normal.
18. Select one of the following run modes to determine where to
enter run information and how to analyze data (see Notes 22
and 23):
500
Shuyi Ma et al.