Next UltraExpress™ FS DNA Library Prep Kit |NEB酶试剂 New England Biolabs

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产品信息

Download the NEBNext UltraExpress FS DNA Library Prep Kit Data Supplement.

The NEBNext UltraExpress FS DNA Library Prep Kit has been developed in response to user need for a faster, streamlined DNA prep workflow. This kit integrates enzymatic fragmentation and delivers high quality libraries from a variety of sample types. It features a single protocol for all DNA inputs, ranging from 10 – 200 ng of intact DNA. The workflow incorporates master mixed reagents, reduced incubation times, and fewer cleanup steps. Use of this kit also generates less plastic consumable waste because the entire library prep is conducted in a single tube. 

  • Go from intact sample to library in under 2 hours with FS (Fragmentation System) enzymatic fragmentation
  • Single-protocol simplicity cuts down on reaction setup time, while streamlined workflows speed up library prep
  • A single-tube workflow means less plastic and consumable waste 
  • Flexibility is enabled with simple guidelines for customized protocols, if desired
  • Automation-friendly protocols for enhanced scalability
Figure 1: NEBNext UltraExpress FS DNA Library Prep workflow

Next UltraExpress™ FS DNA Library Prep Kit |

Figure 2: The NEBNext UltraExpress FS DNA Library Prep Kit provides robust library yields over a wide input range

Next UltraExpress™ FS DNA Library Prep Kit |

Libraries were prepared in triplicate from 10, 50, 100 and 200 ng of a 9:1 Human NA19240 genomic DNA (Coriell Institute for Medical Research) and Escherichia coli gDNA (Lofstrand Labs Limited) mixed sample, using the NEBNext UltraExpress FS DNA single-protocol workflow (e.g., same adaptor amount and 6 PCR cycles for all input amounts). Yields exceeded the minimum requirement (40 ng) for a single Illumina® NovaSeq® 6000 run to achieve whole genome sequencing with at least 30X coverage.

Figure 3: The NEBNext UltraExpress FS DNA Library Prep Kit produces libraries with uniform GC coverage and insert size from a range of input amounts

Next UltraExpress™ FS DNA Library Prep Kit |

Libraries were prepared from 10, 50, 100 and 200 ng of a 9:1 Human NA19240 genomic DNA (Coriell Institute for Medical Research) and Escherichia coli gDNA (Lofstrand Labs Limited) mixed sample, using the NEBNext UltraExpress FS DNA single-protocol workflow (e.g., same adaptor amount and 6 PCR cycles for all input amounts). Libraries were pooled and sequenced on an Illumina NextSeq® 500/550 (2 x 75 bases). Data showed consistent (A) GC coverage and (B) insert size. 2 million paired-end reads from each library were sampled (seqtk v1.0), adaptor-trimmed (seqprep v0.1) and mapped to a composite reference containing GRCh38 and E. coli MG1655 contigs (bowtie2 v2.5.0). GC coverage and insert size distributions were calculated using Picard’s CollectGCBiasMetrics and Picard CollectInsertSizeMetrics (v1.56.0); Picard CollectGCBiasMetrics. (1.56) was run on human autosomes only due to the even copy number assumption of the tool. In (A), the horizontal grey line indicates the expected normalized coverage of 1.0, and the dots in shades of green represent read numbers at each GC%. The grey area plot is a histogram representing the distribution of GC content in 100 bp windows of the reference genome.

Figure 4: The NEBNext UltraExpress FS DNA Library Prep Kit produces representative GC coverage and insert size peaks for microbial genomic DNA over a broad range of GC composition

Next UltraExpress™ FS DNA Library Prep Kit |

Libraries were prepared using the NEBNext UltraExpress FS DNA protocol for 10 ng and 100 ng of genomic DNA from Haemophilus influenzae, Escherichia coli, Rhodopseudomonas palustris and Bordetella pertussis. Data showed (A) representative GC coverage and (B) insert size peaks across samples with genome GC contents of 38%-68% GC. Libraries were pooled and sequenced on an Illumina NextSeq 500/550 (2 x 75 bases). 2 million paired-end reads from each library were sampled (seqtk v1.0), adaptor-trimmed (seqprep v0.1), and aligned to their respective reference genomes (bowtie2 v.2.5.0). GC coverage and insert size distributions were calculated using Picard’s CollectGCBiasMetrics and Picard CollectInsertSizeMetrics (v1.56.0). The horizontal grey line indicates the expected normalized coverage of 1.0, and the colored dots represent read numbers at each GC%. The grey area plot is a histogram representing the distribution of GC content in 100 bp windows of the reference genome for the 10 and 100 ng inputs.

Figure 5: The NEBNext UltraExpress FS DNA Library Prep Kit provides robust library complexity

Next UltraExpress™ FS DNA Library Prep Kit |

Libraries were prepared using the NEBNext UltraExpress FS DNA protocol from (A) 10 ng and 100 ng of the ZymoBIOMICS® Microbial Community DNA Standard (Zymo Research®, Catalog #D6306), and (B) 10 ng and 100 ng of the ZymoBIOMICS Microbial Community DNA Standard II (Log Distribution) (Zymo Research, Catalog #D6311). Libraries were pooled and sequenced on an Illumina® MiSeq® (2 x 75 bases). 750,000 paired-end reads from each library were sampled (seqtk v1.3), adaptor-trimmed (seqprep v0.1) and aligned to a composite reference genome (bowtie2 v2.4.5). 1,000 bp windows of constituent genomes were counted (bedtools 2.30.0) and compared across replicates and input levels. High correlation was observed between replicates and between inputs.

Figure 6: The NEBNext UltraExpress FS DNA Library Prep Kit generates libraries representative of input DNA

Next UltraExpress™ FS DNA Library Prep Kit |

Libraries were prepared using using the NEBNext UltraExpress FS DNA protocol from 10 ng and 100 ng of the ZymoBIOMICS Microbial Community DNA Standard (Zymo Research #D6306). Libraries were pooled and sequenced on an Illumina MiSeq® (2 x 75 bases). 750,000 paired-end reads from each library were sampled (seqtk v1.3), adaptor-trimmed (seqprep v0.1), and aligned to a composite reference genome (bowtie2 v2.4.5). 1,000 bp windows of constituent genomes were counted (bedtools 2.30.0) and compared across expected and detected composition for both input levels. The detection of specific microbial gDNA was consistent with the expected composition. Expected composition: Cryptococcus neoformans 2%, Saccharomyces cerevisiae 2%, Bacillus subtilis 12%, Escherichia coli 12%, Enterococcus faecalis 12%, Lactobacillus fermentum 12%, Listeria monocytogenes 12%, Pseudomonas aeruginosa 12%, Staphylococcus aureus 12% and Salmonella enterica 12%.

Figure 7: The NEBNext UltraExpress FS DNA Library Prep Kit generates libraries representative of input DNA even with complex mixtures across a log range

Next UltraExpress™ FS DNA Library Prep Kit |

Libraries were prepared using the NEBNext UltraExpress FS DNA protocol from 10 ng and 100 ng of the ZymoBIOMICS Microbial Community DNA Standard II (Log Distribution) (Zymo Research, Catalog # D6311). Libraries were pooled and sequenced on an Illumina MiSeq (2 x 75 bases). 750,000 paired-end reads from each library were sampled (seqtk v1.3), adaptor-trimmed (seqprep v0.1), and aligned to a composite reference genome (bowtie2 v2.4.5). 1,000 bp windows of constituent genomes were counted (bedtools 2.30.0) and compared across expected and detected composition for both input levels. Consistent correlation between expected and detected composition was noted (R2 = 0.96 for 10 ng and R2 = 0.97 for 100 ng). Expected composition: Listeria monocytogenes 89.1%, Pseudomonas aeruginosa 8.9%, Bacillus subtilis 0.89%, Saccharomyces cerevisiae 0.89%, Escherichia coli 0.089%, Salmonella enterica 0.089%, Lactobacillus fermentum 0.0089%, Enterococcus faecalis 0.00089%, Cryptococcus neoformans 0.00089% and Staphylococcus aureus 0.000089%.

 

产品类别:
Library Preparation for Illumina Products,
DNA Library Prep for Illumina,
Next Generation Sequencing Library Preparation Products

  • 试剂盒组成

    本产品提供以下试剂或组分：

    NEB # 名称 组分货号 储存温度 数量 浓度
    • E3340S     -20    
        NEBNext UltraExpress FS Enzyme Mix E3343AVIAL -20 1 x 0.024 ml Not Applicable
        NEBNext UltraExpress FS Reaction Buffer E3344AVIAL -20 1 x 0.096 ml Not Applicable
        NEBNext UltraExpress Ligation Master Mix E3345AVIAL -20 1 x 0.48 ml Not Applicable
        NEBNext MSTC High Yield Master Mix E3346AVIAL -20 1 x 1.08 ml Not Applicable
        TE Buffer (1X) E3347AVIAL -20 1 x 0.72 ml 1 X
        NEBNext Bead Reconstitution Buffer E3348AVIAL -20 1 x 1.92 ml Not Applicable
    • E3340L     -20    
        NEBNext UltraExpress FS Enzyme Mix E3343AAVIAL -20 1 x 0.096 ml Not Applicable
        NEBNext UltraExpress FS Reaction Buffer E3344AAVIAL -20 1 x 0.384 ml Not Applicable
        NEBNext UltraExpress Ligation Master Mix E3345AAVIAL -20 2 x 0.96 ml Not Applicable
        NEBNext MSTC High Yield Master Mix E3346AAVIAL -20 1 x 4.32 ml Not Applicable
        TE Buffer (1X) E3347AAVIAL -20 1 x 2.88 ml 1 X
        NEBNext Bead Reconstitution Buffer E3348AAVIAL -20 1 x 7.68 ml 1 X

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操作说明、说明书 & 用法

  • 操作说明

    1. Where can I find protocols for using the NEBNext UltraExpress™ FS DNA Library Prep Kit NEB #E3340)?

  • 说明书

    产品说明书包含产品使用的详细信息、产品配方和质控分析。

    • manualE3340

工具 & 资源

  • Web 工具

    • NEBNext Selector

FAQs & 问题解决指南

  • FAQs

    1. When preparing samples using the NEBNext UltraExpress™ FS DNA Library Prep Kit, can my input DNA be in EDTA-containing solutions?
    2. What if I see a precipitate in the NEBNext UltraExpress™ FS Reaction Buffer?
    3. Do I really need to vortex the NEBNext UltraExpress™ FS Enzyme Mix?
    4. The ligation reaction is very viscous. What will happen if it is not mixed properly?
    5. How much starting material must I use when preparing libraries with the NEBNext UltraExpress™ FS DNA Library Prep Kit?
    6. What sample types can I use with the NEBNext UltraExpress™ FS DNA Library Prep Kit?
    7. Can I use the NEBNext UltraExpress™ FS DNA Library Prep Kit for bisulfite conversion and EM-seq™ workflows?
    8. Is it possible to use sheared DNA as input material with the NEBNext UltraExpress™ FS DNA Library Prep Kit?
    9. What is the difference between the NEBNext UltraExpress™ FS DNA Library Prep Kit and the NEBNext® Ultra™ II FS DNA Library Prep Kit? How do I choose the right one?
    10. Do the NEBNext UltraExpress™ Library Prep kits come with beads?
    11. Which NEBNext® Multiplex Oligos (Adaptors & Primers) can be used with the NEBNext UltraExpress Library Prep Kits?
    12. What should I do if I see adaptor dimer in my NEBNext UltraExpress™ libraries?
    13. Can this kit be used with adaptors and primers from suppliers other than NEB?
    14. Can I use the NEBNext UltraExpress™ FS DNA Library Prep Kit to prepare libraries for sequencing on Ion Torrent instruments?
    15. Does the fragmentation reagent in NEBNext UltraExpress™ FS DNA kits contain NEBNext dsDNA Fragmentase®?
    16. Can the NEBNext UltraExpress™ FS DNA Library Prep Kit provide sufficient library yields for target enrichment?