ZN991 RNA-Seq
dUTP method was used. Strand specificity is maintained by the incorporation of deoxy-UTP during the second strand cDNA synthesis and subsequent destruction of the uridine-containing strand during subsequent step of library construction. First-strand cDNA synthesis reaction was incubated at 25*C for 10 min, followed by 42*C for 1 h and inactivated by incubation at 70*C for 15 min. Double strand cDNAs were repaired with the DNA Terminator End Repair Kit. The Illumina PE Adapter was ligated to the ends of DNA fragments with the Quick Ligation Kit. Ligation product of 250-300 bp fragments was excised. The recovered fragment was digested at 37*C for 15 min with 2 units of Uracil-N- Glycosylase. The digested product was purified on a QIAQuick PCR column followed by 14 cycles of amplification using AccuPrime™ Pfx DNA Polymerase