NEBNext ULTRAEXPRESS® FS DNA Library Prep Kit

Catalog #	Concentration	Size	List Price	Quantity	Your Price
E3340L		96 reactions	$3,290.00		$2,961.00
E3340S		24 reactions	$863.00		$776.70

Catalog #	Size	List Price	Your Price
E3340L	96 reactions	$3,290.00	$2,961.00
E3340S	24 reactions	$863.00	$776.70

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

E3340 Library Yield — 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.

E3340 Input Amounts — 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.

E3340 Sample Types — 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.

E3340 Robust Library Complexity — 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.

e3340 Representative Library Input — 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%.

E3340 Complex Mixtures — 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%.

Reagents Supplied

The following reagents are supplied with this product:

NEB #

Component Name

Component #

Stored at (°C)

Amount

Concentration

E3340S -20

NEBNext UltraExpress FS Enzyme Mix	E3343AVIAL	-20	1 x 0.024 ml
NEBNext UltraExpress^® FS Reaction Buffer	E3344AVIAL	-20	1 x 0.096 ml
NEBNext UltraExpress^® Ligation Master Mix	E3345AVIAL	-20	1 x 0.48 ml
NEBNext MSTC High Yield Master Mix	E3346AVIAL	-20	1 x 1.08 ml
TE Buffer (1X)	E3347AVIAL	-20	1 x 0.72 ml	1 X
NEBNext^® Bead Reconstitution Buffer	E3348AVIAL	-20	1 x 1.92 ml

E3340L -20

NEBNext UltraExpress® FS Enzyme Mix	E3343AAVIAL	-20	1 x 0.096 ml
NEBNext UltraExpress^® FS Reaction Buffer	E3344AAVIAL	-20	1 x 0.384 ml
NEBNext UltraExpress^® Ligation Master Mix	E3345AAVIAL	-20	2 x 0.96 ml
NEBNext MSTC High Yield Master Mix	E3346AAVIAL	-20	1 x 4.32 ml
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

Properties & Usage

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FAQs

Quality Control Assay

Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

Specifications

The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

E3340S_L_G_v1

Certificate of Analysis

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

Safety Data Sheets

NEBNext UltraExpress^® FS DNA Library Prep Kit

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