NEBNext DNA Overview

Illumina

SOLiD

Roche/454

Ion Torrent

For DNA library preparation for sequencing with the 454 platform, reagents from NEB are available in a set, master mix or module format. Choose from reagents for the new quick fragment library prep (end repair, dA-tailing, adaptor ligation and size selection) or the original method (end repair, adaptor ligation fill-in and ssDNA isolation). Each of these reagents has been functionally validated by preparation of a genomic DNA library followed by 454 sequencing™

• Next generation sequencing sample preparation
  
• High density hybridization arrays
• Genomic subtraction hybridization methods
• Expression library construction

• Convenient formats –All of the required enzymes, buffers and nucleotides are included, and are available in set or master mix format. Modules offer the ability to customize sample preparation method.
• Flexibility – Choose from reagents and master mixes for either the quick or original fragment library preparation.
• Functional Validation – Each reagent set or module is functionally validated by preparation of a genomic DNA library followed by 454 sequencing.
• Compatible with all instruments – Each set or module can be used with GS FLX Titanium and GS Junior.
• Stringent Quality Controls – Additional QCs ensure maximum quality and purity.
• Value pricing

Workflow for DNA Sample Prep	NEBNext Quick DNA Library Prep Reagent Set for 454	NEBNext Quick DNA Library Prep Master Mix Set for 454
End Repair/dA Tailing	T4 DNA Polymerase T4 PNK Taq DNA Polymerase dNTPs ATP NEBuffer 2 (10X)	End Prep Enzyme Mix End Repair Reaction Buffer (10X) Nuclease-Free Water
Adaptor Ligation	Quick T4 DNA Ligase	Quick T4 DNA Ligase
Size Selection	Sizing Buffer (1.2X) TE	Sizing Buffer (1.2X) TE

Workflow for DNA Sample Prep	NEBNext DNA Library Prep Reagent Set for 454	NEBNext DNA Library Prep Master Mix Set for 454	NEBNext Fill-in and ssDNA Isolation Module
End Repair	T4 DNA Polymerase T4 PNK dNTPs ATP BSA (10X) NEBuffer 2 (10X)	End Repair Enzyme Mix End Repair Reaction Buffer (10X) Nuclease-Free Water
Adaptor Ligation	Quick T4 DNA Ligase Quick Ligation Reaction Buffer (2X)	Quick T4 DNA Ligase Quick Ligation Reaction Buffer (5X)
Fill-in and ssDNA Isolation	Bst DNA Polymerase, Large Fragment ThermoPol Reaction Buffer (10X) dNTPs Hydrophilic Magnetic Streptavidin Magnetic Beads Bead Binding Buffer (2X) Bead Wash Buffer (1X)	Bst DNA Polymerase, Large Fragment Adapter Fill-In Reaction Buffer (10X) Hydrophilic Magnetic Streptavidin Magnetic Beads Bead Binding Buffer (2X) Bead Wash Buffer (1X)	Bst DNA Polymerase, Large Fragment Adapter Fill-In Reaction Buffer (10X) Hydrophilic Magnetic Streptavidin Magnetic Beads Bead Binding Buffer (2X) Bead Wash Buffer (1X)

		Quick Videos to Help Guide your Next Gen Sequencing Work: Next Gen Sequencing: Master Mix or Aliquot? A Breakthrough Method of RNA Sample Prep Common Mistakes to Avoid in Next Gen Sample Prep