FAQ: Can I perform poly(A) mRNA isolation or rRNA depletion and then freeze my samples at -80°C before beginning library preparation with the NEBNext UltraExpress™ RNA Library Prep Kit?

Yes, however, some modifications will be required. If you plan to freeze your mRNA- or rRNA-depleted RNA before library prep, elution of your samples in 4 µl is recommended. 2.5µl of this eluate will then be transferred into a fresh tube and combined with an equivalent volume (2.5 µl) of NEBNext UltraExpress RNA Fragmentation Mix., for a total volume of 5 µl. Slightly greater sample loss is expected using this method compared to the standard protocol. However, adaptor input and cycling recommendations will remain the same.