Single Cell Gene Expression Protocol v3.1 | Loading Chromium Next GEM Chip G

Combine the master mix with water and single cell suspension, then keep the mixture on ice until ready to load.

Ensure the Chromium Next GEM single cell 3 Prime V 3.1 gel beads have been equilibrated to room temperature for at least 30 minutes and prepare the partitioning oil for loading.

Gently pipette mix the master mix and cell suspension, then dispense 70 microliters into the chip row labeled 1, controlling the plunger rate to approximately 5 seconds per stroke.

Snap the gel bead tube strip into a 10x Vortex adapter, vortex for 30 seconds, then centrifuge for 5 seconds and verify no bubbles are present.

Puncture the foil seal and slowly aspirate 50 microliters of gel beads, then dispense into the row labeled 2 without introducing air bubbles and wait 30 seconds.

Dispense 45 microliters of partitioning oil from a reagent reservoir into the wells in row labeled 3 to ensure proper GEM generation.

Close the secondary holder lid, carefully handle the loaded chip without tipping or pressing the gasket, and insert it into the Chromium instrument for processing.