Prepare and Add Custom Primers

1.

If frozen, thaw each custom primer to be used.

2.

Use NovaSeq X Series Custom Primer Buffer to dilute custom primers to yield the following volumes at 0.3 µM final concentration. When combining primers to make the custom Index Read or any custom read primer mixture, each primer should be at 0.3 µM final concentration.

Custom Primer	Volume (ml) for 1.5B Flow Cell	Volume (ml) for 10B or 25B Flow Cell
Custom Read 1 primer	3.0	5.0
Custom Read 2 primer	3.0	5.0
Custom Index 1 and 2 primer mix	5.0	6.0

1.

Use a polynit heatseal wipe to wipe clean the foil seal covering each custom primer position.

Position	Custom Primer
CP1	Custom Read 1 primer
CP2	Custom Read 2 primer
CP3	Custom Index 1 and 2 primer mix

2.

Using a clean pipette tip, pierce the foil seal covering each custom primer position.

3.

Add the following volumes of custom primer to the appropriate position on the reagent cartridge. Avoid touching the foil seal as you dispense the primer.

Position	Custom Primer	Volume (ml) for 1.5B Flow Cell	Volume (ml) for 10B or 25B Flow Cell
CP1	Custom Read 1 primer	3.0	5.0
CP2	Custom Read 2 primer	3.0	5.0
CP3	Custom Index 1 and 2 primer mix	5.0	6.0