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New England Biolabs

NEB NEBNext Ultra II FS DNA PCR-Free Library Prep Kit (Cat No. E7430)

NEB NEBNext Ultra II FS DNA PCR-Free Library Prep Kit (Cat No. E7430)

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Don’t be biased! The NEBNext Ultra II FS DNA PCR-free Library Prep Kit for Illumina offers an amplification-free workflow for DNA-seq based on the streamlined and reliable Ultra II FS workflow. Starting with as little as 50 ng of DNA, get the high-quality, high-yield libraries that you need without PCR bias.

  • Includes NEBNext FS (Fragmentation System) enzymatic fragmentation reagents
  • Perform fragmentation, end repair and dA-tailing with a single enzyme mix 
  • 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 50 ng to 500 ng intact DNA
  • 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 3′ single T overhang

For convenience, a version of this kit is available with SPRIselect beads.

Note: For sheared DNA, we recommend the NEBNext Ultra II DNA PCR-free Library Prep Kit for Illumina.

 

Catalog # Concentration Size
E7430S Not Applicable 24 reactions
E7430L Not Applicable 96 reactions

 

 

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

Advantages

  • Enjoy flexible input amounts from 50 ng to 500 ng of intact DNA
  • Experience reliable fragmentation with a single protocol, regardless of DNA input amount or GC content 
  • Perform fragmentation, end repair and dA-tailing with a single enzyme mix 
  • 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) (NEB #E7395) or other Illumina-compatible index adaptors with a 3’ single T overhang

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

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



A. PCR-free libraries were prepared with human cell line NA19240 genomic DNA (Corielle Institute) using NEBNext Ultra II FS DNA PCR-free and Illumina PCR-free library prep kits and size selected for 350 bp long inserts.

B. Libraries of 150-200 bp inserts were prepared using the enzyme-based fragmentation kits NEBNext Ultra II FS DNA PCR-free and Roche Sequencing Kapa HyperPlus PCR-free library prep kits without size selection. NEBNext Unique Dual Index UMI Adaptors DNA Set 1 and Kapa Dual-Indexed Adaptors were used for the NEBNext and Kapa kits, respectively, following manufacturers' recommendations. 



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




NEBNext Ultra II FS DNA PCR-free libraries have more even genome coverage than Illumina DNA PCR-free. Sequencing libraries were prepared according to the manufacturer's recommendations from 250 ng of NA19240 cell line DNA by 2 different users 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 references (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 assigned 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. There is significant variation between users for Illumina PCR-free libraries; however, the FS DNA PCR-free libraries are very consistent between users.



 

 

 

 

 

Product Name: NEB NEBNext Ultra II FS DNA PCR-Free Library Prep Kit (Cat No. E7430)

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