User's Manual

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Optional: Set up flow cytometry with Cell sorter control kit (Cat. No. CO10100)
To easily set up flow cytometry of dPCR droplets, use the Samplix Cell sorter control kit (Cat. No. CO10100).
This kit consists of ready-made dPCR droplets with a defined and large population of positive droplets. The
kit allows you to establish the settings for dPCR droplet sorting.
1. Make sure the 2x dPCR buffer ● is diluted with molecular grade water to 1x. Mix well by vortexing
for 10 seconds or inverting the tube at least 20 times.
2. Stain the Control droplets ○ with Droplet dye ● as follows:
• Prepare flow cytometry buffer by adding 500 μl 1x dPCR-buffer to a flow cytometry
tube (tubes depend on flow cytometer instrument).
• Spin down the vial of Droplet dye ● at 5000 rpm for 2 minutes.
• Add 5 μl Droplet dye ● into the flow cytometry tube with dPCR buffer. Mix gently to
dissolve the dye in the dPCR-buffer.
• Transfer 50 μl of Control droplets ○ to the prepared flow cytometry tube. Use tips
that minimize binding of droplets to the side of the tip.
• Leave mix of droplets and dPCR buffer at room temperature protected from light
for 5 min to stain droplets.
3. Load the tube on a flow cytometer and start the analysis.
4. Identify the dPCR droplets as described above (see Fig. 4.1).
5. Identify positive and negative fluorescent droplets as described above (see Fig. 4.2). In the Cell
sorter control droplets, you can expect the fraction of positive droplets to be about 10-15% of the
total observed dPCR droplets.