Isolate cDNA from PIPs

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• CEB
• 0.2 ml 8-tube strips and caps
1. Label one new tube strip per sample.
1. If you are resuming after a safe stopping point, briefly centrifuge the tubes.
2. Add 40 µl CEB to each cDNA reaction in the 8-tube strips. Seal with a new cap.
3. Pulse vortex, and then centrifuge for 5 seconds.
4. Place into the guide rack, and then tap the rack against the bench three times.
5. Let sit until PIPs have settled below the guide wire (~1 minute).
6. Without disturbing the pellet, transfer 60 µl supernatant to a new tube strip.
7. Add 60 μl CEB to the 0.2 ml tube containing the PIP pellet.
8. Pulse vortex, and then centrifuge for 5 seconds.
9. Place into the guide rack, and then tap the rack against the bench three times.
10. Let sit until PIPs have settled below the guide wire (~1 minute).
11. Without disturbing the pellet, transfer 60 μl supernatant from the tube containing the PIP pellet to the tube containing the transferred supernatant.

The total volume of supernatant per sample is 120 μl.

12. Briefly centrifuge the tube containing the supernatant, and then examine the bottom of the tubes for PIPs.
13. If residual PIPs pellets are present, transfer the supernatant to a new tube strip, and then measure the transferred supernatant volume.

If the final volume of supernatant is not 120 μl per sample, adjust the magnetic bead volume during magnetic bead purification.

14. Save the remaining PIP pellet and store at -85°C to -65°C for up to 3 months.

The saved pellets are required for Illumina Single Cell Supplemental Enrichment and Amplification Kit applications. If library preparation fails to produce sequencing-quality libraries, the saved pellets can be used to recover cDNA.