NEBNext® Multiplex Oligos Selection Chart

 

SINGLE INDEX

DUAL INDEX

UNIQUE DUAL INDEX

UNIQUE DUAL INDEX UMIs

NEBNext Multiplex Oligos for Illumina®

Contains UMI

No

No

No

Yes

Addresses index hopping

No

No

Yes

Yes

Indexing Strategy

Index Primer

Index Primer

Index Primer

Index Adaptor

Adaptor Included
NEBNext Adaptor (loop)  NEBNext Adaptor (loop) Standard: NEBNext Adaptor (loop)
LV: None; adaptor provided with library prep kit
Unique Dual Index UMI Adaptor

Applications

DNA-seq, RNA-seq 
(except Small RNA)

DNA-seq, RNA-seq
(except Small RNA)

DNA-seq, RNA-seq
(except Small RNA)

PCR-free DNA-seq, RNA-seq 
(except Small RNA)

Number of indices  for multiplexing

up to 144

up to 384

up to 480

384 for DNA; 96 for RNA

Compatible with ARTIC sequencing for Illumina
Yes Yes  Yes No

Compatible with EM-seq

Yes*

Yes*

Yes*

No

Compatible with EpiMark® bisulfite sequencing

Yes**

Yes**

Yes**

No

Number of sets available; formats and indices available

Five; Sets 1-4 (12 indices/set): Individual vials 96 Index: Premixed plate

Two; individual vials containing 8 i5 primers and 12 i7 primers for combinatorial mixing

Thirteen; Sets are provided as either 24 or 96 indices in premixed, foil-sealed 96-well plates; Including a version for EM-seq (up to 120 indices, either 96-well plate or 24 vial format) 

Four sets for DNA, One set for RNA; Adaptors with 96 indices in premixed, foil-sealed 96-well plate (DNA-seq or RNA-seq) and primers

* Requires use of the correct adaptor; choose based on the version of EM-seq being used. Original EM-seq (NEB #E7120) is not supplied with the NEBNext EM-seq Adaptor. The adaptor and indices must be purchased separately as part of NEBNext Unique Dual Index Pairs for EM-seq (NEB #E7140). Adaptors for use with NEBNext EM-seq v2 (NEB #E8015) and NEBNext E5hmC-seq (NEB #E3350) are provided as components in the library prep kits. Index primers for use with these kits are optimized for low-volume workflows and are available as NEBNext LV Unique Dual Index Primers (Sets 1-5).

**  Requires use of NEBNext EM-seq adaptor from NEBNext Multiplex Oligos for Enzymatic Methyl-seq (Unique Dual Index Primer Pairs, #E7140S/L) .