Single cell library preparation

Good afternoon-

1) BSA is an effective surfactant that suppresses clumping and improves pelleting without damaging cells.

2) 2kxg for 2 minutes. It may be necessary to remove 900ul and spin again.

3) Heat shock response would be swamped by osmotic & cell wall stress response.

4) RNAlater is saturated ammonium sulfate and will precipitate RNA in place, freezing the transcriptome and preventing changes during processing.

5) Yes; a small amount of background RNA is expected, but can be quantified after sequencing. We see fewer than 10 counts per barcode in non-cell droplets, which is a combination of cells that lyse prior to encapsulation, template switching, and sequencing error.

Thanks

-Chris

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