G4™ Sequencing Platform Quick Reference Card
| Loading Concentration|
10 pM 15 pM 20 pM 25 pM 30 pM
Thaw reagent cartridges at 2°C to 8°C for 2 days and equilibrate to room temperature for 30 minutes, or thaw in a room temperature water bath for 2.5 hours (wipe off residual water).
Remove flow cell from storage and equilibrate to room temperature for 30 minutes.
Download a sample data sheet template.
Fill out the Header, Settings, and Data sections.
Save the complete sample data sheet in the SampleData folder or in a location available to the instrument.
Make fresh 0.2 M NaOH by mixing 800 µL molecular-grade water with 200 µL 1 M NaOH and invert tube several times to mix.
Dilute the optional Singular PhiX control to 150 pM with TE. Vortex and then centrifuge briefly.
Denature the library and PhiX control in a separate new tubes. Vortex and then centrifuge briefly. For variations, see the Loading Concentration Calculator.
Incubate for 5 minutes at room temperature.
To the same tubes, add Tris-HCl to neutralize, then vortex to mix and centrifuge briefly.
Place the librarylibraries on ice until ready to proceed.
Dilute the 200 pM library and PhiX control in Sample Loading Buffer to the loading concentration in a new tube. Vortex and then centrifuge briefly. Loading concentration may differ for your application.
Keep on ice until ready to use, no longer than 2 hours.
Obtain a sample cartridge for each flow cell.
Equilibrate the sample cartridge to room temperature for 30 minutes.
Remove the sample cartridge from the sealed bag. Avoid extended light exposure.
Pipette 260 µL diluted library into the appropriate well associated with each lane as listed in the sample data sheet.
Set aside until prompted to load sample cartridges onto the G4 instrument.
Empty the waste reservoir.
Invert reagent cartridges 5–10 times, then gently tap the cartridge on the bench surface.
Remove each flow cell from the packaging and remove the seals covering the ports and any seal adhesive.
When prompted load the wash buffer cartridges, flow cells, reagent cartridges, and sample cartridges.
After the pre-run system check is complete, select Start Run.