A Low-Cost Library Construction Protocol and Data Analysis Pipeline for Illumina-Based Strand-Specific Multiplex RNA-Seq
Figure 2
Enforcing strand specificity using dUTP.
dTTP is substituted with dUTP during second strand cDNA synthesis. Y-shaped (partial-complementary) adapters are ligated and the dUTP-marked strand is digested with uracil-DNA gylcosylase (UDG). PCR amplification of this single strand confers strand specificity.