SEQanswers - View Single Post - Single cell RNA-seq trouble

alekzs · 01-05-2018, 02:07 PM

I have tried increasing the PCR cycles up to 23 but still got this double-bump pattern.
The samples were frozen, I sort directly into lysis buffer (max 5min per plate), directly spin the plate down and then freeze on dry-ice and transfer to -80C until processing. For processing, I start out with bringing the RNA beads to room temp, RNAzap the work area, let the plates thaw on ice for 10min and then start the protocol.

01-05-2018, 02:07 PM	#4
alekzs Junior Member Location: CT Join Date: Jan 2018 Posts: 8	I have tried increasing the PCR cycles up to 23 but still got this double-bump pattern. The samples were frozen, I sort directly into lysis buffer (max 5min per plate), directly spin the plate down and then freeze on dry-ice and transfer to -80C until processing. For processing, I start out with bringing the RNA beads to room temp, RNAzap the work area, let the plates thaw on ice for 10min and then start the protocol.