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

omans · 01-05-2018, 01:40 PM

Primary lymphocytes have a significantly lower level of mRNA than cultured cells.

I've found that scRNA-seq is very sensitive to sample handling. Are you using fresh or frozen T cells? How long is your sort?

Also, how many PCR cycles did you perform for each of these samples? For other lymphocyte types, I've needed to exceed 21 cycles to see a peak.

01-05-2018, 01:40 PM	#3
omans Junior Member Location: New York Join Date: Jan 2018 Posts: 2	Primary lymphocytes have a significantly lower level of mRNA than cultured cells. I've found that scRNA-seq is very sensitive to sample handling. Are you using fresh or frozen T cells? How long is your sort? Also, how many PCR cycles did you perform for each of these samples? For other lymphocyte types, I've needed to exceed 21 cycles to see a peak.