NEBNext® DNA Library Prep Master Mix Set for 454™


The NEBNext® DNA Library Prep Master Mix Set for 454™ contains enzymes and buffers in convenient master mix formulations that are ideally suited for sample preparation for next-generation sequencing, and for preparation of single stranded DNA for use in high density hybridization arrays or for genomic subtraction hybridization methods. Each of these components must pass rigorous quality control standards and are lot controlled, both individually and as a set of reagents.

Lot Control

The lots provided are managed separately and qualified by additional functional validation. Individual reagents undergo standard enzyme activity and quality control assays, and also meet stringent criteria in the additional quality controls listed on each individual component page

Reagents Supplied

The following reagents are supplied with this product:

Store at (°C)Concentration
NEBNext End Repair Enzyme Mix
NEBNext End Repair Reaction Buffer10X
Quick T4 DNA Ligase-20
NEBNext Quick Ligation Reaction Buffer-205X
Bst DNA Polymerase, Large Fragment
NEBNext Adapter Fill-in Reaction Buffer10X
Molecular Biology Grade Water
Hydrophilic Streptavidin Magnetic Beads4 mg/ml
NEBNext Bead Wash Buffer1X
NEBNext Bead Binding Buffer2X

Properties and Usage

Storage Notes

  • -20°C for box 1, 4°C for box 2



    • NEBNext End Repair Enzyme Mix
    • NEBNext End Repair Reaction Buffer (10X)
    • Quick T4 DNA Ligase
    • NEBNext Quick Ligation Reaction Buffer (5X)
    • Bst DNA Polymerase, Large Fragment
    • NEBNext Adaptor Fill-in Reaction Buffer (10X) 
    • Molecular Biology Grade Water


    • Hydrophilic Streptavidin Magnetic Beads (4 mg/ml)
    • NEBNext Bead Binding Buffer (2X)
    • NEBNext Bead Wash Buffer (1X)
  2. Quality control values for a specific lot can be found on the datacard which accompanies each product.


  1. Maricic, T. and S. Paabo (2009). Optimization of 454 sequencing library preparation from small amounts of DNA permits sequence determination of both DNA strands.  Biotechniques. 46, 51-52, 54-57.
  2. Straus, D. and F.M. Ausubel (1990). Proc. Natl. Acad. Sci. USA. . 87, 1889-1893.
  3. Zhou, X. and D.T. Wong (2007). Methods Mol. Biol. . 396, 295-314.


  1. NEBNext DNA Sample Prep Master Mix Set for 454 End Repair Module Protocol (E6070)
  2. NEBNext DNA Sample Prep Master Mix Set for 454 Quick Ligation Module Protocol (E6070)
  3. NEBNext DNA Sample Prep Master Mix Set for 454 Fill-in and ssDNA Isolation Module Protocol (E6070, E6071)


The Product Manual includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name these document files: manual[Catalog Number].

Interactive Tools


  • Kuleshov, K. et al. (2014). Whole-genome haplotyping using long reads and statistical methods. Nature Biotechnology. doi:10.1038/nbt.2833. PubMedID: 24561555

Safety Data Sheet

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.