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

Confirm that the single cell suspension and master mix are on ice, and verify that gel beads have been equilibrated to room temperature for at least 30 minutes before proceeding with chip assembly.

Ensure the secondary holder is closed, then attach the gasket with the notch in the upper left corner by hooking it onto the left tabs, pulling toward the right to engage the right tabs, and avoiding contact with the smooth gasket surface.

Remove the chip from the sealed bag and insert it into the secondary holder by aligning the notches, sliding the chip left under the guide, and depressing the right side until the spring-loaded clip engages.

Add 70 microliters of 50% glycerol into the unused wells in row one by gently touching the pipette tip to the bottom surface and slowly withdrawing as you dispense.

Add 50 microliters of 50% glycerol into the unused wells in row two using the same pipetting technique of gently touching the tip to the bottom and slowly withdrawing.

Add 45 microliters of 50% glycerol into the unused wells in row three, ensuring that no glycerol is added to the bottom row labeled 'no fill' and that no substitutes are used.

After glycerol loading is complete, prepare the master mix with water and single cell suspension, then load into the designated chip wells for the experiment.