Protocol for use with NEBNext ULTRAEXPRESS® RNA Library Prep Kit (NEB#E3330) and NEBNext rRNA Depletion Kits (NEB #E7400, #E7405) and non indexed adaptor v1

The NEBNext UltraExpress RNA Library Prep Kit contains the enzymes and buffers required to to rapidly convert 25–250 ng of total RNA into high-quality libraries for next-generation sequencing on the Illumina platform. The fast, simple workflow features minimal hands-on time and allows use of a single adaptor dilution and PCR cycling condition across the entire input range. In addition to the standard protocol, an appendix is included that details customized adaptor and cycling recommendations for varying RNA input amounts, if further optimization for library yields is required. If a stopping point is required between mRNA enrichment/ribosomal RNA depletion and library preparation, please refer to the FAQs tab for considerations and recommendations the FAQ tab is on NEB.com.