Fig. 8
From: Advancing brain barriers RNA sequencing: guidelines from experimental design to publication
Library preparation protocols. Commonly used library preparation protocols for RNA sequencing. For RNA-Seq, a first step of ribosomal RNA (rRNA) depletion from the total RNA is performed; random primers are used for reverse transcription; dUTPs are used for the second strand synthesis; Y-shaped adaptors are then ligated, and then the second strand containing dUTPs is depleted, allowing to retrieve stranded information. 3′ RNA-Seq makes use of oligo dT primers for the reverse transcription, which selects mRNA. Using template switching, the second adapter is incorporated in the cDNA molecules. Small RNA-Seq uses adaptors that ligate to the small RNAs and allow the reverse transcription