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NEBNext® Ultra™ II DNA PCR-free Library Prep Kit for Illumina®

Catalog # Concentration Size List Price Quantity Your Price
E7410L 96 reactions $3,184.00
$2,865.60
E7410S 24 reactions $837.00
$753.30
Catalog # Size List Price Your Price
E7410L 96 reactions $3,184.00
$2,865.60
E7410S 24 reactions $837.00
$753.30
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

Don’t be biased! The NEBNext Ultra II DNA PCR-free Library Prep Kit for Illumina offers an amplification-free workflow for DNA-seq based on the streamlined and reliable Ultra II workflow. Starting with as little as 250 ng of DNA, get the high-quality, high-yield libraries that you need without PCR bias.

  • Save time with an optimized PCR-free workflow, reduced hands-on time and automation compatibility
  • Generate high quality libraries without an amplification step using inputs from 250 ng to 1,000 ng
  • Benefit from improved library uniformity
  • Use with the NEBNext Multiplex Oligos for Illumina (Unique Dual Index UMI Adaptors DNA Set 1) or other Illumina-compatible index adaptors with a 3’ single T overhang

Also available with SPRIselect beads.

Note: If your input DNA is not sheared, we recommend the NEBNext Ultra II FS DNA PCR-free Library Prep Kit for Illumina, which fragments, end repairs, and dA-tails DNA, all in a single tube with no cleanups or transfers

 

The NEBNext Ultra II DNA PCR-free Library Prep Kit contains the enzymes and buffers required to convert a broad range of sheared DNA input amounts into libraries for next-generation sequencing on the Illumina platform, all without an amplification step. The fast, user-friendly workflow requires minimal hands-on time, resulting in excellent library yields and GC coverage without requiring PCR. 

Advantages

  • Enjoy flexible input amounts from 250 ng to 1,000 ng of sheared DNA
  • Get more of what you need with reliably high library yields
  • Prepare PCR-free libraries with a broad range of sample types 
  • Save time with streamlined workflows, reduced hands-on time, and automation compatibility
  • Use with the NEBNext Multiplex Oligos for Illumina (Unique Dual Index UMI Adaptors DNA Set 1) or other Illumina-compatible index adaptors with a 3’ single T overhang

A separate version of the NEBNext Ultra II DNA PCR-free Library Prep Kit is available with sample purification beads.

Figure 1: NEBNext Ultra II DNA PCR-free Library Prep Kit generates libraries with higher yields

A. PCR-free libraries were prepared with NA19240 genomic DNA (Coriell Institute) using NEBNext Ultra II DNA PCR-free and Illumina TruSeq® PCR-free library prep kits and size selected for 350 bp inserts. DNA inputs were 1 µg.

B. Libraries of 150-200 bp inserts were prepared using NEBNext Ultra II DNA PCR-free and Roche Sequencing Kapa HyperPrep library prep kits coupled with Covaris shearing without size selection. NEBNext Unique Dual Index UMI Adaptors DNA Set 1, IDT for Illumina (TruSeq DNA UD Indexes) and Kapa Dual-Indexed Adaptors were used for the NEBNext, Illumina, and Kapa kits, respectively, following manufacturers' recommendations.



Figure 2: NEBNext Ultra II DNA PCR-free Library Prep provides uniform genome coverage with human DNA


NEBNext Ultra II DNA PCR-free libraries have even more genome coverage than Illumina DNA PCR-free libraries. Sequencing libraries were prepared according to the manufacturer’s recommendations from 250 ng of NA19240 cell line DNA and sequenced on a single NovaSeq 6000 S4 v1.5 flow cell (PE 2x150 bp). Reads were adaptor trimmed (fastp 0.20.0), aligned to the Telomere-to-Telomere chm13 reference (draft 1, bwa-mem 0.7.17), and duplicate marked (samblaster 0.1.24). 2.8B reads were randomly sampled from each bam (sambamba view -s 0.6.8) Coverage of primary, pass-filter assignments was assessed using mosdepth (v0.3.1, -F 772) and plotted for autosomes or the mitochondrial genome (inset). Expected coverage (num_reads*read length/genome size) is indicated).

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  NEBNext® Ultra II End Prep Enzyme Mix E7646AVIAL -20 1 x 0.072 ml
  NEBNext® Ultra II End Prep Reaction Buffer E7647AVIAL -20 1 x 0.168 ml
  NEBNext® Ligation Enhancer E7374AVIAL -20 1 x 0.024 ml
  NEBNext® Ultra™ II Ligation Master Mix E7648AVIAL -20 1 x 0.72 ml
  NEBNext® Ultra II End Prep Enzyme Mix E7646AAVIAL -20 1 x 0.288 ml
  NEBNext® Ultra II End Prep Reaction Buffer E7647AAVIAL -20 1 x 0.672 ml
  NEBNext® Ligation Enhancer E7374AAVIAL -20 1 x 0.096 ml
  NEBNext® Ultra™ II Ligation Master Mix E7648AAVIAL -20 3 x 0.96 ml
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