Hi,
I'm staring to learn about single cell transcriptome sequencing, about smart-seq2(Sandberg 2014https://www.nature.com/articles/nprot.2014.006.pdf), after the first strand synthesis with TSO, then the ISPCR primers were added for cDNA amplification. But I don't really understand why there seems 2 of the ISPCR primers anneal to the first strand at the same time, does that mean the complementary strand can be generated from 2 directions. I'm so confused... If anyone could kindly explain that to me, I'd be much appreciated.
I'm staring to learn about single cell transcriptome sequencing, about smart-seq2(Sandberg 2014https://www.nature.com/articles/nprot.2014.006.pdf), after the first strand synthesis with TSO, then the ISPCR primers were added for cDNA amplification. But I don't really understand why there seems 2 of the ISPCR primers anneal to the first strand at the same time, does that mean the complementary strand can be generated from 2 directions. I'm so confused... If anyone could kindly explain that to me, I'd be much appreciated.