NEBNext DNA Library PreparationProduct information
Library preparation for the major next generation sequencing platforms requires the ligation of specific adaptor oligos to fragments of the DNA to be sequenced. First, DNA is fragmented to the optimal length determined by the downstream platform. Because DNA fragmentation does not result in homogeneous, blunt-ended fragments, end repair is needed to ensure that each molecule is free of overhangs, and contains 5’ phosphate and 3’ hydroxyl groups.
Libraries to be used in blunt-ended adaptor ligation, including Ion Torrent™ library construction, can be used directly in the ligation step. For Illumina® libraries and some libraries intended for the 454™ platform, incorporation of a non-templated deoxyadenosine 5’-monophosphate (dAMP) onto the 3’ end of blunted DNA fragments, a process known as dA-tailing, is necessary. dA-tails prevent concatamer formation during downstream ligation steps, and enable DNA fragments to be ligated to adaptors with complementary dT-overhangs.
The desired adaptor ligated DNA size for Illumina, and Ion Torrent platforms can be selected via gel electrophoresis before amplification by the polymerase chain reaction (PCR). New England Biolabs supplies reagents for DNA library preparation for the leading sequencing platforms. For more information, choose the DNA Library Construction Workflow tab below.
Advantages of NEBNext Library Prep Reagents:
- Convenient formats – All of the required enzymes, buffers and nucleotides are included, many available in master mix format. Sets for Ion Torrent include adaptors and primers. Modules offer the ability to customize sample preparation.
- Functional Validation – Each reagent set or module is functionally validated by preparation of a genomic DNA or ChIP-Seq DNA library that is sequenced using the appropriate sequencing platform (Illumina, 454, or Ion Torrent).
- Stringent Quality Controls – Additional QCs ensure maximum quality and purity.
- Value Pricing
Applications of NEBNext Library Prep Reagents:
- Next generation sequencing library preparation
- Expression library construction
- High density hybridization arrays
- Genomic subtraction hybridization methods
NEBNext® dsDNA Fragmentase® is an enzyme-based solution for fragmentation of DNA. dsDNA Fragmentase will generate 100-800 bp fragments in a time-dependent manner, and is ideal for generation of fragments for library construction and next generation sequencing sample preparation.
Advantages of NEBNext dsDNA Fragmentase:
- High throughput compatibility
- Control of fragment size
- Generates 100-300 base pair fragments more effectively than nebulization
- Enzyme-based fragmentation results in cleaner ends as compared to mechanical shearing
- Value pricing
NEBNext offers a solution for each step of the DNA Library Construction Workflow
*Also available as a set of individual reagents. See ordering information.
** The DNA sample Prep set for SOLiD was discontinued on 02/06/2014
|LIBRARY PREP SETS||NEB #||SIZE|
|NEBNext Ultra DNA Library Prep Kit for Illumina||E7370S/L||24/96 reactions|
|NEBNext DNA Library Prep Master Mix Set for Illumina||E6040S/L||12/60 reactions|
|NEBNext ChIP-Seq Library Prep Master Mix Set for Illumina||E6240S/L||12/60 reactions|
|NEBNext DNA Library Prep Reagent Set for Illumina||E6000S/L||12/60 reactions|
|NEBNext ChIP-Seq Library Prep Reagent Set for Illumina||E6200S/L||12/60 reactions|
|NEBNext Fast DNA Fragmentation & Library Prep Set for Ion Torrent||E6285S/L||10/50 reactions|
|NEBNext Fast DNA Library Prep Set for Ion Torrent||E6270S/L||10/50 reactions|
|NEBNext Quick DNA Library Prep Master Mix Set for 454||E6090S/L||10/50 reactions|
|NEBNext DNA Library Prep Master Mix Set for 454||E6070S/L||10/50 reactions|
|NEBNext Quick DNA Library Prep Reagent Set for 454||E6080S/L||10/50 reactions|
* This product was discontinued on 02/06/2014
|MODULES & ENZYMES||NEB #||SIZE||COMPATIBLE
|NEBNext End Repair Module||E6050S/L||20/100 reactions||Multiple|
|NEBNext dA-Tailing Module||E6053S/L||20/100 reactions||Illumina|
|NEBNext Quick Ligation Module||E6056S/L||20/100 reactions||Multiple|
|NEBNext Fill-in and ssDNA Isolation Module||E6071S/L||20/100 reactions||454|
|NEBNext Singleplex Oligos for Illumina||E7350S/L||12/60 reactions||Illumina|
|NEBNext Multiplex Oligos for Illumina (Index Primers Set 1)||E7335S/L||24/96 reactions||Illumina|
|NEBNext Multiplex Oligos for Illumina (Index Primers Set 2)||E7500S/L||24/96 reactions||Illumina|
|NEBNext dsDNA Fragmentase||M0348S/L||50/250 reactions||Multiple|
|NEBNext High-Fidelity 2X PCR Master Mix||M0541S/L||50/250 reactions||Multiple|
|NEBNext dsDNA Fragmentase Reaction Buffer||B0348S||6 x 1 ml||Multiple|
|NEBNext End Repair Reaction Buffer||B6052S||2 ml||Multiple|
|NEBNext Quick Ligation Reaction Buffer||B6058S||2 ml||Multiple|
|NEBNext dA-Tailing Reaction Buffer||B6059S||2 ml||Multiple|
454™ is a trademark of Roche.
ILLUMINA® is a registered trademark of Illumina, Inc.
ION TORRENT™ and SOLID™ are trademarks owned by Life Technologies, Inc.
NEBNEXT® and FRAGMENTASE® are registered trademarks of New England Biolabs, Inc..
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