Load Chromium Next GEM Chip K:
a. Assemble Chromium Next GEM Chip
Attach the gasket by holding the tongue (curved end, to the right) and hook the gasket on the left-hand tabs of the holder.
Gently pull the gasket toward the right and hook it on the two right-hand tabs.
Remove the chip from the sealed bag. Use the chip within ≤ 24 h
b. Dispense 50% Glycerol Solution into each unused wells
(if processing <8 samples/chip)
i. 70 µl in each unused well in row labeled 1
ii. 50 µl in each unused well in row labeled 2
iii.45 µl in each unused well in row labeled 3
c. Prepare Master Mix + Cell Suspension
Refer to the Cell Suspension Volume Calculator Table
Add the appropriate volume of nuclease-free water to Master Mix.
Add the appropriate volume of single cell suspension to Master Mix
(Total of 75 µl in each tube)
Gently pipette mix the single cell suspension before adding to the Master Mix.
Gently pipette mix the Master Mix + Cell Suspension
Using the same pipette tip, dispense 70 µl Master Mix + Cell Suspension into the bottom center of each well in row labeled 1 without introducing bubbles.
• Snap the tube strip holder with the Gel Bead strip into a 10x Vortex Adapter. Vortex 30 sec.
• Centrifuge the Gel Bead strip for ~5 sec.
• Confirm there are no bubbles at the bottom of the tubes and the liquid levels are even.
• Place the Gel Bead strip back in the holder. Secure the holder lid
• Puncture the foil seal of the Gel Bead tubes.
• Slowly aspirate 50 µl Gel Beads.
• Dispense into the wells in row labeled 2 without introducing bubbles.
• Dispense 45 µl Partitioning Oil into the wells in row labeled 3 from a reagent reservoir. Failure to add Partitioning Oil to the top row labeled 3 will prevent GEM generation and can damage the Chromium Controller
Close the lid (gasket already attached). DO NOT touch the smooth side of the gasket. DO NOT press down on the top of the gasket.
Run the chip in the Chromium Controller or X/iX immediately after loading the Partitioning Oil.