NEBNext^® LV Unique Dual Index Primers Set 3

The following reagents are supplied with this product:

NEB #	Component Name	Component #	Stored at (°C)	Amount	Concentration
NEB # Stored at (°C) E3404S -20 Component Name Component # Stored at (°C) Amount Concentration NEBNext® LV Unique Dual Index Primers Set 3 E3404AVIAL -20 1 x 5 μl/well 10 µM

Companion Products

• NEBNext^® Enzymatic 5hmC-seq Kit
• NEBNext^® LV Unique Dual Index Primers (Unique Dual Index Set 2B)
• NEBNext^® Enzymatic Methyl-seq v2 Kit
• NEBNext^® Enzymatic Methyl-seq v2 Conversion Module
• NEBNext^® LV Unique Dual Index Primers Set 2A
• NEBNext^® LV Unique Dual Index Primers Set 1

1. Where can I find protocols for use of the NEBNext® LV Unique Dual Index Primers Set 3, including index pooling guidelines? (NEB #E3404)

• Forward Strand Workflow Sample Sheet for E3404: NovaSeq 6000 with v1.0 reagent kits, MiniSeq with Rapid reagent kits, MiSeq, HiSeq 2000/2500 (paired-end flow cell), HiSeq 3000/4000 (single-read flow cell)
• NEB_E3404_UDI_IlluminaLRM
• Reverse Complement Workflow Sample Sheet for E3404: iSeq 100, MiniSeq with Standard reagent kits, NextSeq Systems, NovaSeq 6000 with v1.5 reagent kits, HiSeq 2000/2500 (single-read flow cell), HiSeq 3000/4000 (paired-end flow cell)

• NEBNext® Multiplex Oligos Selection Chart

• NEBNext Selector

1. What is the difference between NEBNext® LV Unique Dual Index Primers and NEBNext LV Unique Dual Index Primers?
2. Are Sample Sheets available for use with the NEBNext® LV Unique Dual Index Primers Sets 1-5?
3. Are libraries that have been dual indexed using the NEBNext LV Unique Dual Index Primers compatible with single-end sequencing
4. How many primer pairs are available in the NEBNext® LV Unique Dual Index Primer Pairs Sets 1-5?
5. Can the NEBNext® LV Unique Dual Index Primers be used to address “index hopping” with Illumina® patterned flow cells?
6. Are the NEBNext® LV Unique Dual Index Primers the same as NEBNext Multiplex Oligos for Illumina®?
7. Are the NEBNext® LV Unique Dual Index Primers validated for the EM-seq™ v2 or E5hmC-seq™ workflows?
8. What is the concentration of the Index Primers in NEBNext® LV Unique Dual Index Primers?
9. How many reactions’ worth of primer are in each well of the NEBNext® LV Unique Dual Index Primers?
10. Can I use the 96-well primer plate supplied with the NEBNext LV Unique Dual Index Primers to do my PCR reactions
11. Do I need to spike in custom sequencing primers when sequencing libraries made with NEBNext® LV Unique Dual Index Primers on Illumina® sequencing instruments?
12. Can I perform single read runs and still get both index sequences? Can I run both single read and paired-end recipes with dual-indexed libraries?
13. What sequences need to be trimmed for NEBNext libraries that are sequenced on an Illumina instrument?
14. What do I do if I received an index collision/barcode collision error?

Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

• E3404S_v2

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

• E3404S_v1_10222070
• E3404S_v2_10292833
• E3404S_v2_10304543
• E3404S_v2_10269511
• E3404S_v2_10320958

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This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.