New England Biolabs, E7771S, NEBNext® Ultra™ II RNA First Strand Synthesis Module

Related Categories Automation for NEBNext® NGS Library Prep,, RNA Library Prep for Illumina,, Next Generation Sequencing Library Preparation Specification Materials Required but not Supplied NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB #E7490) or NEBNext® rRNA Depletion Kit v2 (Human/Mouse/Rat) (NEB #E7400 or NEB #E7405) or NEBNext® Globin & rRNA Depletion Kit (Human/Mouse/Rat) (NEB #E7750 or NEB #E7755) or NEBNext® rRNA Depletion Kit (Bacteria) (NEB #E7850 or NEB #E7860) or NEBNext® RNA Depletion Core Reagent Set (NEB #E7865 or NEB #E7870) or NEBNext rRNA Depletion Kit (Human/Mouse/Rat) (NEB #E6310 or NEB #E6350) NEBNext Ultra II Directional RNA Second Strand Synthesis Module (NEB #E7550) or NEBNext Ultra II Non-Directional RNA Second Strand Synthesis Module (NEB #E6111) Magnetic rack or plate (e.g., NEBNext® Magnetic Separation Rack (NEB #S1515S), Alpaqua ® 96S Super Magnet Plate (#A001322), or equivalent) FAQ Q: What other materials do I need in addition to the NEBNext Ultra II First Strand Synthesis Module for RNA-seq experiments? A: If you are starting from total RNA, we recommend either the NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB #E7490) or the NEBNext rRNA Depletion Kit v2 (Human/Mouse/Rat) (NEB #E7400 or NEB #7405) or NEBNext® Globin & rRNA Depletion Kit (Human/Mouse/Rat) (NEB #E7750 or NEB #E7755) or NEBNext® rRNA Depletion Kit (Bacteria) (NEB #E7850 or NEB #E7860) or NEBNext® RNA Depletion Core Reagent Set (NEB #E7865 or NEB #E7870) or NEBNext rRNA Depletion Kit (Human/Mouse/Rat) (NEB #E6310 or NEB #E6350) for use prior to first strand synthesis. For strand-specific RNA-seq workflows, use the following kits: NEBNext Ultra II Directional RNA Second Strand Synthesis Module (NEB #E7550) NEBNext Ultra II End Repair/dA-Tailing Module (NEB #E7546) NEBNext Ultra II Ligation Module (NEB #E7595) NEBNext Ultra II Q5 Master Mix (NEB #M0544) For non-directional RNA-seq workflows, use the following kits: NEBNext Ultra II Non-Directional RNA Second Strand Synthesis Module (NEB #E6111) NEBNext Ultra II End Repair/dA-Tailing Module (NEB #E7546) NEBNext Ultra II Ligation Module (NEB #E7595) NEBNext Ultra II Q5 Master Mix (NEB #M0544) For adaptors and primers, NEBNext Oligos are compatible, as are adaptors and primers from other sources. Q: What if I have already fragmented my total RNA and need to perform the First Strand Synthesis Reaction? A: If you are starting from fragmented total RNA, we recommend using the NEBNext rRNA Depletion Kit (Human/Mouse/Rat; #E6310) prior to first strand synthesis. Follow the protocol in Chapter 3 of the Ultra II Directional RNA Library Prep Kit (#E7760) for strand-specific RNA-seq workflows. For non-directional RNA-seq workflows, follow the protocol in Chapter 3 of the Ultra II RNA Library Prep Kit (#E7770). Q: What if I have already fragmented my mRNA and just need to perform the First Strand Synthesis Reaction? A: If you are starting from fragmented mRNA, we recommend following the protocol in Chapter 5 of the Ultra II Directional RNA Library Prep Kit (#E7760) for strand-specific RNA-seq workflows. For non-directional RNA-seq workflows, follow the protocol in Chapter 5 of the Ultra II RNA Library Prep Kit (#E7770). Q: What is the composition of the random primers? A: The random primers are random hexamers supplied at a 1.9 mM.