Home Protocols Where can I find protocols for using NEBNext® RNA Library Prep reagents?

Where can I find protocols for using NEBNext® RNA Library Prep reagents?

Use the table below to find the appropriate protocol for your library prep, based on sample type and kit being used.

NEBNext ADAPTOR (NON-INDEXED)

NEBNext Ultra II Directional
(NEB#E7760,E7765)		 NEBNext Ultra II Non-Directional
(NEB#E7770,E7775)		 No Library Prep
Intact + Partially Fragmented Fragmented or FFPE Intact + Partially Fragmented Fragmented or FFPE
No Enrichment Protocol Section 4 Protocol Section 5 Protocol Section 4 Protocol Section 5 N/A
Poly(A) mRNA Magnetic Isolation Module (NEB#E7490) Protocol Section 1 N/A Protocol Section 1 N/A Manual NEBNext Poly(A) mRNA Magnetic Isolation Module
Legacy rRNA Depletion (Human/ Mouse/ Rat) (NEB#E6310,E6350) Protocol Section 2 Protocol Section 3 Protocol Section 4 Protocol Section 5 Protocol Section 1
rRNA Depletion Kit v2 (Human/ Mouse/ Rat) (NEB#E7400,E7405) Protocol Section 2 Protocol Section 3 Protocol Section 2 Protocol Section 3 Protocol Section 1
Globin and rRNA Depletion Kit (Human/ Mouse/ Rat) (NEB#E7750,E7755) Protocol Section 2 N/A Protocol Section 3 N/A Protocol Section 1
rRNA Depletion Kit Bacteria (NEB#E7850,E7860) Protocol Section 2 Protocol Section 3 Protocol Section 4 Protocol Section 5 Protocol Section 1
RNA Depletion Core Reagent Set (NEB#E7865,E7870) Protocol Section 2 Protocol Section 3 Protocol Section 4 Protocol Section 5 Protocol Section 1
Globin and rRNA Depletion Kit + Poly(A) mRNA Magnetic Isolation Module (NEB#E7750orE7755andE7490) Protocol Section 4 N/A N/A N/A N/A

 

UNIQUE DUAL INDEX UMI ADAPTOR*

NEBNext Ultra II Directional
(NEB#E7760,E7765)		 NEBNext Ultra II Non-Directional
(NEB#E7770,E7775)
Intact + Partially Fragmented Fragmented or FFPE Intact + Partially Fragmented Fragmented or FFPE
No Enrichment Protocol Section 4 Protocol Section 5 Protocol Section 4 Protocol Section 5
Poly(A) mRNA Magnetic Isolation Module (NEB#E7490) Protocol Section 1 N/A Protocol Section 1 N/A
Legacy rRNA Depletion (Human/ Mouse/ Rat) (NEB#E6310,E6350) Protocol Section 2 Protocol Section 3 Protocol Section 2 Protocol Section 3
RNA Depletion Kit v2 (Human/ Mouse/ Rat) (NEB#E7400,E7405) Protocol Section 1 Protocol Section 2 Protocol Section 3 Protocol Section 4
Globin and rRNA Depletion Kit (Human/ Mouse/ Rat) (NEB#E7750,E7755) Protocol Section 1 N/A Protocol Section 2 N/A
rRNA Depletion Kit Bacteria (NEB#E7850,E7860) Protocol Section 1 Protocol Section 2 Protocol Section 3 Protocol Section 4
RNA Depletion Core Reagent Set (NEB#E7865,E7870) N/A N/A N/A N/A
Globin and rRNA Depletion Kit + Poly(A) mRNA Magnetic Isolation Module (NEB#E7750orE7755andE7490) N/A N/A N/A N/A

* Please also see manual for NEBNext® Multiplex Oligos for Illumina® (Unique Dual Index UMI Adaptors RNA Set 1)

 

Protocols for using NEBNext Ultra II RNA Library Prep Kits for Illumina, NEBNext Poly(A) mRNA Magnetic Isolation Module, NEBNext RNA Depletion modules and NEBNext Multiplex Oligos can be found within the following manuals. Please refer to the following details to identify which manual contains the protocol relevant to your product combination.

 

Click here for protocols for NEBNext rRNA Depletion Kit (Human/Mouse/Rat) (NEB #E6310, #E6350S), including:
  • Protocol for Depletion of Intact or Partially Degraded RNA using NEBNext rRNA Depletion Kit (Human/Mouse/Rat) (NEB #E6310, #E6350)
  • Protocol for Library Prep of Intact or Partially Degraded RNA using NEBNext rRNA Depletion Kit (Human/Mouse/Rat) (NEB #E6310, #E6350) and NEBNext Ultra II Directional RNA Library Prep Kit  (NEB #E7760, #E7765); with non -indexed adaptors
  • Protocol for Library Prep of Degraded RNA (e.g., FFPE) using NEBNext rRNA Depletion Kit (Human/Mouse/Rat) (NEB #E6310, #E6350) and NEBNext Ultra II Directional RNA Library Prep Kit (NEB #E7760, #E7765); with non -indexed adaptors
  • Protocol for Library Prep of Intact or Partially Degraded RNA using NEBNext rRNA Depletion Kit (Human/Mouse/Rat) (NEB #E6310, #E6350) and NEBNext Ultra II Non- Directional RNA Library Prep Kit for Illumina (NEB #E7770, #E7775); with non -indexed adaptors
  • Protocol for Library Prep of Degraded RNA (e.g., FFPE) using NEBNext rRNA Depletion Kit (Human/Mouse/Rat) (NEB #E6310, #E6350) and NEBNext Ultra II Non- Directional RNA Library Prep Kit (NEB #E7770, #E7775); with non -indexed adaptors

 

 

Click here for the protocol for NEBNext rRNA Depletion Kit v2 (Human/Mouse/Rat) NEB #E7400, #E7405), including:

  • Protocol for Depletion of Intact or Partially Degraded RNA using NEBNext rRNA Depletion Kit v2 (Human/Mouse/Rat) (NEB #E7400, #E7405)

 

Click here for protocols for use with NEBNext Ultra II Directional RNA Library Prep Kit NEBNext (NEB #E7760, #E7765), including:

  • Protocol for Library Prep of Intact or Partially Degraded RNA using NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB #E7490) and NEBNext Ultra II Directional RNA Library Prep Kit (NEB #E7760, #E7765); with non -indexed adaptors
  • Protocol for Library Prep of Intact or Partially Degraded RNA using NEBNext rRNA Depletion Kit v2 (Human/Mouse/Rat) (NEB #E7400, #E7405) and NEBNext Ultra II Directional RNA Library Prep Kit (NEB #E7760, #E7765); with non -indexed adaptors
  • Protocol for Library Prep of Degraded RNA (e.g., FFPE) using NEBNext rRNA Depletion Kit v2 (Human/Mouse/Rat) (NEB #E7400, #E7405) and NEBNext Ultra II Directional RNA Library Prep Kit (NEB #E7760, #E7765); with non -indexed adaptors
  • Protocol for Library Prep of Purified mRNA or rRNA Depleted RNA (or other intact or partially degraded RNA not requiring depletion) using NEBNext Ultra II Directional RNA Library Prep Kit (NEB #E7760, #E7765); with non -indexed adaptors
  • Protocol for Library Prep of Degraded RNA (e.g., FFPE or other degraded RNA not requiring depletion) using NEBNext Ultra II Directional RNA Library Prep Kit (NEB #E7760, #E7765); with non -indexed adaptors

 

Click here for protocols for use with NEBNext Ultra II Non-Directional RNA Library Prep Kit (NEB #E7770, #E7775), including:  
  • Protocol for Library prep of Intact or Partially Degraded RNA using NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB #E7490) and NEBNext Ultra II Non-Directional RNA Library Prep Kit (NEB #E7770, #E7775); with non -indexed adaptors
  • Protocol for Library prep of Intact or Partially Degraded RNA using NEBNext rRNA Depletion Kit v2 (Human/Mouse/Rat) (NEB #E7400, #E7405) and NEBNext Ultra II Non-Directional RNA Library Prep Kit (NEB #E7770, #E7775); with non -indexed adaptors
  • Protocol for Library Prep of Degraded RNA (e.g., FFPE) using NEBNext rRNA Depletion Kit v2 (Human/Mouse/Rat) (NEB #E7400, #E7405) and NEBNext Ultra II Non- Directional RNA Library Prep Kit (NEB #E7770, #E7775); with non -indexed adaptors
  • Protocol for Library Prep of Purified mRNA or rRNA Depleted RNA (or other intact or partially degraded RNA not requiring depletion) using NEBNext Ultra II Non-Directional RNA Library Prep Kit (NEB #E7770, #E7775); with non -indexed adaptors
  • Protocol for Library Prep of Degraded RNA (e.g., FFPE or other degraded RNA not requiring depletion) using NEBNext Ultra II Non- Directional RNA Library Prep Kit (NEB #E7770, #E7775); with non -indexed adaptors

 

Click here for protocols for use with NEBNext Globin and rRNA Depletion Kit (Human/Mouse/Rat) (NEB #E7750, #E7755), including:  
  • Protocol for Depletion of Intact or Partially Degraded RNA using NEBNext Globin and rRNA Depletion Kit (Human/Mouse/Rat) (NEB #E7750, E7755)
  • Protocol for Library Prep of Intact or Partially Degraded RNA using NEBNext Globin and rRNA Depletion Kit (Human/Mouse/Rat) (NEB #E7750, E7755), NEBNext Ultra II Directional RNA Library Prep Kit (NEB #E7760, #E7765); with non -indexed adaptors
  • Protocol for Library Prep of Intact or Partially Degraded RNA using NEBNext Globin and rRNA Depletion Kit (Human/Mouse/Rat) (NEB #E7750, E7755), NEBNext Ultra II Non-Directional RNA Library Prep Kit (NEB #E7770, #E7775); with non -indexed adaptors
  • Protocol for Enrichment of mRNAs, Excluding Globin mRNA, from Whole Blood Total RNA using the Globin & rRNA Depletion with NEBNext Globin & rRNA Depletion Kit (Human/Mouse/Rat) (NEB #E7750, #E7755), NEBNext Poly(A) mRNA Magnetic Isolation Module NEB #E7490), NEBNext Ultra II Directional RNA Library Prep Kit (NEB # 7760, #E7765); with non -indexed adaptors

 

Click here for protocols for use with NEBNext rRNA Depletion Kit (Bacteria) (NEB #E7850, NEB #E7860), including: 
  • Protocol for Depletion of Intact or Partially Degraded RNA using NEBNext rRNA Depletion Kit (Bacteria) (NEB #E7850, NEB #E7860)
  • Protocol for Library Prep of Intact or Partially degraded RNA using NEBNext rRNA Depletion Kit (Bacteria) (NEB #E7850, NEB #E7860) and NEBNext Ultra II Directional RNA Library Prep Kit  (NEB #E7760, NEB #E7765); with non -indexed adaptors
  • Protocol for Library Prep of Degraded RNA using NEBNext rRNA Depletion Kit (Bacteria) (NEB #E7850, NEB #E7860) and NEBNext Ultra II Directional RNA Library Prep Kit (NEB #E7760, NEB #E7765); with non -indexed adaptors
  • Protocol for Library Prep of Intact or Partially Degraded RNA using NEBNext rRNA Depletion Kit (Bacteria) (NEB #E7850, NEB #E7860) and NEBNext Ultra II Non- Directional RNA Library Prep Kit (NEB #E7770, NEB #E7775); with non -indexed adaptors
  • Protocol for Library Prep of Degraded RNA using NEBNext rRNA Depletion Kit (Bacteria) (NEB #E7850, NEB #E7860) and NEBNext Ultra II Non- Directional RNA Library Prep Kit (NEB #E7770, NEB #E7775); with non -indexed adaptors

 

Click here for protocols for use with NEBNext RNA Depletion Core Reagent Set (NEB #E7865, NEB #E7870), including:  
  • Protocol for Depletion of Intact or Partially Degraded RNA using NEBNext RNA Depletion Core Reagent Set (NEB #E7865, NEB #E7870)
  • Protocol for Library Prep of Intact or Partially Degraded RNA using NEBNext RNA Depletion Core Reagent Set (NEB #E7865, NEB #E7870) and NEBNext Ultra II Directional RNA Library Prep Kit  (NEB #E7760, NEB #E7765); with non -indexed adaptors
  • Protocol for Library Prep of Degraded RNA (e.g., FFPE) using NEBNext RNA Depletion Core Reagent Set (NEB #E7865, NEB #E7870) and NEBNext Ultra II Directional RNA Library Prep Kit (NEB#E7760, NEB#E7765); with non -indexed adaptors
  • Protocol for Library Prep of Intact or Partially Degraded RNA using NEBNext RNA Depletion Core Reagent Set (NEB #E7865, NEB #E7870) and NEBNext Ultra II Non- Directional RNA Library Prep Kit  (NEB #E7770, NEB #E7775); with non -indexed adaptors
  • Protocol for Library Prep of Degraded RNA (e.g., FFPE) using NEBNext RNA Depletion Core Reagent Set (NEB #E7865, NEB #E7870) and NEBNext Ultra II Non- Directional RNA Library Prep Kit  (NEB #E7770, NEB #E7775); with non -indexed adaptors

 

Click here for the protocols for NEBNext rRNA Depletion Kit v2 (Human/Mouse/Rat) (NEB #E7400, #E7405) Indexed, UMI adaptor; including:
  • Protocol for Library Prep of Intact or Partially Degraded RNA using NEBNext rRNA Depletion Kit v2 (Human/Mouse/Rat) (NEB #E7400, #E7405) and NEBNext Ultra II Directional RNA Library Prep Kit (NEB #E7760, #E7765); with indexed, UMI adaptors
  • Protocol for Library Prep of Degraded RNA (e.g., FFPE) using NEBNext rRNA Depletion Kit v2 (Human/Mouse/Rat) (NEB #E7400, #E7405) and NEBNext Ultra II Directional RNA Library Prep Kit (NEB #E7760, #E7765); with indexed, UMI adaptors
  • Protocol for Library Prep of Intact or Partially Degraded RNA using NEBNext rRNA Depletion Kit v2 (Human/Mouse/Rat) (NEB #E7400, #E7405) and NEBNext Ultra II Non- Directional RNA Library Prep Kit (NEB #E7770, #E7775); with indexed, UMI adaptors
  • Protocol for Library Prep of Degraded RNA (e.g., FFPE) using NEBNext rRNA Depletion Kit v2 (Human/Mouse/Rat) (NEB #E7400, #E7405) and NEBNext Ultra II Non- Directional RNA Library Prep Kit for Illumina (NEB #E7770, #E7775); with indexed, UMI adaptors

 

Click here for the protocol for NEBNext Multiplex Oligos for Illumina (Unique Dual Index UMI Adaptors RNA Set 1 (NEB #E7416), including: 
  • Setting up the PCR Reactions with NEBNext Multiplex Oligos for Illumina (Unique Dual Index UMI Adaptors RNA)
  • Index Pooling Guidelines for NEBNext Multiplex Oligos for Illumina (Unique Dual Index UMI Adaptors RNA) (NEB #E7416)

 

Click here for protocols for use with NEBNext Globin and rRNA Depletion Kit (Human/Mouse/Rat) (NEB #E7750, E7755), Indexed, UMI adaptor; including:
  • Protocol for Library Prep of Intact or Partially Degraded RNA using NEBNext Globin and rRNA Depletion Kit (Human/Mouse/Rat) (NEB #E7750, E7755) and NEBNext Ultra II Directional RNA Library Prep Kit (NEB #E7760, #E7765); with indexed, UMI adaptors
  • Protocol for Library Prep of Intact or Partially Degraded RNA using NEBNext Globin and rRNA Depletion Kit (Human/Mouse/Rat) (NEB #E7750, E7755) and NEBNext Ultra II Non-Directional RNA Library Prep Kit (NEB #E7770, #E7775); with indexed, UMI adaptors

 

Click here for protocols for use with NEBNext Ultra II Directional RNA Library Prep Kit NEBNext (NEB #E7760, #E7765), Indexed, UMI adaptor; including: 
  • Protocol for Library Prep of Intact or Partially Degraded RNA using NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB #E7490) and NEBNext Ultra II Directional RNA Library Prep Kit (NEB #E7770, #E7775); with indexed, UMI adaptors
  • Protocol for Library Prep of Intact or Partially Degraded RNA using NEBNext rRNA Depletion Kit v2 (Human/Mouse/Rat) (NEB #E7400, #E7405) and NEBNext Ultra II Directional RNA Library Prep Kit (NEB #E7760, #E7765); with indexed, UMI adaptors
  • Protocol for Library Prep of Degraded RNA (e.g., FFPE) using NEBNext rRNA Depletion Kit v2 (Human/Mouse/Rat) (NEB #E7400, #E7405) and NEBNext Ultra II Directional RNA Library Prep Kit (NEB #E7760, #E7765); with indexed, UMI adaptors
  • Protocol for Library Prep of Purified mRNA or rRNA Depleted RNA (or other intact or partially degraded RNA not requiring depletion) using NEBNext Ultra II Directional RNA Library Prep Kit (NEB #E7760, #E7765); with indexed, UMI adaptors
  • Protocol for Library Prep of Degraded RNA (e.g., FFPE or other degraded RNA not requiring depletion) using NEBNext Ultra II Directional RNA Library Prep Kit (NEB #E7760, #E7775); with indexed, UMI adaptors

 

Click here for protocols for use with NEBNext Ultra II Non-Directional RNA Library Prep Kit (NEB #E7770, #E7775), Indexed, UMI adaptor; including: 
  • Protocol for Library Prep of Intact or Partially Degraded RNA using NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB #E7490) and NEBNext Ultra II Non- Directional RNA Library Prep Kit (NEB #E7770, #E7775); with indexed, UMI adaptors
  • Protocol for Library Prep of Intact or Partially Degraded RNA using NEBNext rRNA Depletion Kit v2 (Human/Mouse/Rat) (NEB #E7400, #E7405) and NEBNext Ultra II Non- Directional RNA Library Prep Kit (NEB #E7770, #E7775); with indexed, UMI adaptors
  • Protocol for Library Prep of Degraded RNA (e.g., FFPE) using NEBNext rRNA Depletion Kit v2 (Human/Mouse/Rat) (NEB #E7400, #E7405) and NEBNext Ultra II Non- Directional RNA Library Prep Kit (NEB #E7770, #E7775); with indexed, UMI adaptors
  • Protocol for Library Prep of Purified mRNA or rRNA Depleted RNA (or other intact or partially degraded RNA not requiring depletion) using NEBNext Ultra II Non- Directional RNA Library Prep Kit (NEB #E7770, #E7775); with indexed, UMI adaptors
  • Protocol for Library Prep of Degraded RNA (e.g., FFPE or other degraded RNA not requiring depletion) using NEBNext Ultra II Non- Directional RNA Library Prep Kit (NEB #E7770, #E7775); with indexed, UMI adaptors

 

Click here for protocols for use with NEBNext rRNA Depletion Kit (Bacteria) (NEB #E7850, NEB #E7860), Indexed, UMI adaptor; including: 
  • Protocol for Library Prep of Intact or Partially Degraded RNA using NEBNext rRNA Depletion Kit (Bacteria) (NEB #E7850, NEB #E7860) and NEBNext Ultra II Directional RNA Library Prep Kit (NEB #E7760, NEB #E7765); with indexed, UMI adaptors
  • Protocol for Library Prep of Degraded RNA using NEBNext rRNA Depletion Kit (Bacteria) (NEB #E7850, NEB #E7860) and NEBNext Ultra II Directional RNA Library Prep Kit (NEB #E7760, NEB #E7765); with indexed, UMI adaptors
  • Protocol for Library Prep of Intact or Partially Degraded RNA using NEBNext rRNA Depletion Kit (Bacteria) (NEB #E7850, NEB #E7860) and NEBNext Ultra II Non-Directional RNA Library Prep Kit for Illumina (NEB #E7770, NEB #E7775); with indexed, UMI adaptors
  • Protocol for Library Prep of Degraded RNA using NEBNext rRNA Depletion Kit (Bacteria) (NEB #E7850, NEB #E7860) and NEBNext Ultra II Non- Directional RNA Library Prep Kit (NEB #E7770, NEB #E7775); with indexed, UMI adaptors