Using single nuclei for RNA-seq to capture the transcriptome of postmortem neurons

. Author manuscript; available in PMC: 2016 Jul 12.

Published in final edited form as: Nat Protoc. 2016 Feb 18;11(3):499–524. doi: 10.1038/nprot.2016.015

Smart-Seq2 Procedure step³	Modification	Reason
9: Reverse transcription	TSO, oligo-dT and ISPCR oligos have a 5′ biotin modification	Reduces concatamer formation and increases gene mapping percentage for sequence reads
14: cDNA PCR thermal cycling	Increase the number of amplification cycles from 18 to 21	To compensate for the lower amount of RNA in a nucleus compared with a whole cell