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sequencing
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scg-protocols-v1

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## 1.2 Load GEM-X Chip
- l After removing chip from the sealed bag, use in ≤24 h .
- l Open the lid (gasket attached) of the assembled chip and lay flat for loading.
- l Ensure that the Gel Beads are properly thawed and ready to use.
- l When loading the chip, raising and depressing the pipette plunger should each take ~5 sec . When dispensing, raise the pipette tips at the same rate as the liquid is rising, keeping the tips slightly submerged.
## Color Legend
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- 2: Gel beads
- 1: Sample
3: Oil
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The Chromium X/iX (X Series) Chip Holder, Chip Gasket X/iX, and GEM-X chip images shown below are representative and do not show the specific color &amp; label. Chip holder and gasket should be black and blue in color, respectively. Refer to Assemble GEM-X Chip on page 32 for details.
## a. If loading less than 8 samples/chip, add 50% glycerol solution to each unused well in row 1, 2, and 3
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- l 60 μ l in each unused well in row labeled 1
- l 60 μ l in each unused well in row labeled 2
- l 250 μ l in each unused well in row labeled 3
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DO NOT add 50% glycerol solution to the wells in top row labeled NO FILL.
DO NOT use any substitute for 50% glycerol solution.
## b. Prepare Gel Beads
- l Snap the tube strip holder with the Gel Bead strip into a 10x Vortex Adapter. Vortex 30 sec .
- l Centrifuge the Gel Bead strip for ~5 sec .
- l Confirm there are no bubbles at the bottom of the tubes &amp; the liquid levels are even.
- l Place the Gel Bead strip back in the holder. Secure the holder lid.
GEM-X 3' Chip, gasket attached Representative chip image
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Glycerol in GEM-X 3' Chip
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## c. Prepare Master Mix + Cell suspension
- l Refer to the Cell Suspension Volume Calculator Table on page 34.
- l Add the appropriate volume of PBS to Master Mix.
- l Add corresponding volume of single cell suspension into the Master Mix. Total of 65 μ l in each tube. Gently pipette mix the cell suspension before adding to the Master Mix.
## d. Load Row Labeled 1
- l Gently pipette mix the Master Mix + Cell Suspension.
- l Using the same pipette tip, dispense 60 μ l Master Mix + Cell Suspension into the bottom center of each well in row labeled 1 without introducing bubbles.
- l Wait 30 sec .
## e. Load Row Labeled 2
- l Puncture the foil seal of the Gel Bead tubes.
- l Slowly aspirate 60 μ l Gel Beads.
- l Dispense into the bottom center of each well in row labeled 2 without introducing bubbles.
- l Wait 30 sec .
## f. Load Row Labeled 3
- l Dispense 250 μ l Partitioning Oil B into the wells in row labeled 3 by pipetting two aliquots of 125 μ l from a reagent reservoir.
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