上海金畔生物科技有限公司代理New England Biolabs（NEB）酶试剂全线产品，欢迎访问官网了解更多产品信息和订购。
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
Figure 2: The NEBNext UltraExpress FS DNA Library Prep Kit provides robust library yields over a wide input range
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
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
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
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
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
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
相关产品
相关产品
T2030 Monarch RNA Cleanup Kit 10 ug
NEBNext® 多样本接头引物试剂盒 5（96 种 Unique 双端 Index 引物）
NEBNext® 文库定量试剂盒（Illumina®）
NEBNext® 磁性分离架
操作说明、说明书 & 用法
操作说明
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
When preparing samples using the NEBNext UltraExpress™ FS DNA Library Prep Kit, can my input DNA be in EDTA-containing solutions?
What if I see a precipitate in the NEBNext UltraExpress™ FS Reaction Buffer?
Do I really need to vortex the NEBNext UltraExpress™ FS Enzyme Mix?
The ligation reaction is very viscous. What will happen if it is not mixed properly?
How much starting material must I use when preparing libraries with the NEBNext UltraExpress™ FS DNA Library Prep Kit?
What sample types can I use with the NEBNext UltraExpress™ FS DNA Library Prep Kit?
Can I use the NEBNext UltraExpress™ FS DNA Library Prep Kit for bisulfite conversion and EM-seq™ workflows?
Is it possible to use sheared DNA as input material with the NEBNext UltraExpress™ FS DNA Library Prep Kit?
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?
Do the NEBNext UltraExpress™ Library Prep kits come with beads?
Which NEBNext® Multiplex Oligos (Adaptors & Primers) can be used with the NEBNext UltraExpress Library Prep Kits?
What should I do if I see adaptor dimer in my NEBNext UltraExpress™ libraries?
Can this kit be used with adaptors and primers from suppliers other than NEB?
Can I use the NEBNext UltraExpress™ FS DNA Library Prep Kit to prepare libraries for sequencing on Ion Torrent instruments?
Does the fragmentation reagent in NEBNext UltraExpress™ FS DNA kits contain NEBNext dsDNA Fragmentase®?
Can the NEBNext UltraExpress™ FS DNA Library Prep Kit provide sufficient library yields for target enrichment?
上海金畔生物科技有限公司代理New England Biolabs（NEB）酶试剂全线产品，欢迎访问官网了解更多产品信息和订购。
产品信息
The ARTIC SARS-CoV-2 protocol from the ARTIC Network is a multiplexed amplicon approach covering the whole viral genome. The NEBNext ARTIC SARS-CoV-2 FS Library Prep Kit is based on this method and incorporates optimized and novel reagents.
A single RT protocol is used regardless of input amount (10-10,000 viral genome copies), and no normalization step is required ahead of targeted amplification.
This kit includes two options for primers, including new VarSkip Short v2, for improved Omicron coverage
The new VarSkip (for Variant Skip) Short v2 primers have been designed at NEB to reduce the impact of SARS-CoV-2 variants on amplification and provide improved performance, including with the Omicron variant. VarSkip Short v2 primer pools generate ~550bp amplicons. Please see the FAQ section and the product manual for additional detail. Information on primer variant overlaps can be found here.
The V3 ARTIC primers have been balanced, using methodology developed at NEB based on empirical data from sequencing. In combination with optimized reagents for RT-PCR, the kits deliver improved uniformity of amplicon yields from gRNA across a wide copy number range.
Library preparation uses the NEBNext FS reagents and workflow, incorporating enzymatic cDNA fragmentation, and reagent volumes tailored to the ARTIC application. Note that adaptors and primers for library prep are purchased separately. The novel NEBNext Library PCR Master Mix allows use of the same number of PCR cycles to amplify all libraries, regardless of initial input amounts.
The Express protocol option removes the cleanup of cDNA amplicons and the cleanup of adaptor-ligated DNA. The Express protocol is recommended for inputs of ≥ 100 copies of the (SARS-CoV-2) viral genome.
The NEBNext ARTIC SARS-CoV-2 FS Library Prep Kit generates libraries with inserts ~150 bp. If longer inserts are preferred, the NEBNext ARTIC SARS-CoV-2 Library Prep Kit (NEB E7650) produces ~400 bp insert libraries, compatible with 2 x 250 Illumina sequencing. Note that this kit does not contain VarSkip v2 primers. For Oxford Nanopore Technologies® sequencing the NEBNext ARTIC SARS-CoV-2 Companion Kit (Oxford Nanopore Technologies) (NEB #E7660) can be used. If only the reagents for cDNA synthesis and targeted cDNA amplification are required, the NEBNext ARTIC SARS-CoV-2 RT-PCR Module (NEB #E7626) is available.
Figure 1: Sequence coverage of SARS-CoV-2 genomes with VarSkip Short v2 plus spike-in primers
Consensus genomes known to be generated using VarSkip Short 2 (+ supplementary primers) (green) are compared with other genomes (yellow) submitted to Genbank between 2022-08-01 and 2022-09-24. Genomes were aligned to the NC_045512.2 reference (minimap2 -x map-ont -r 20000 –score-N=0, v.2.24). Ns were tabulated using Qualimap (v2.2.2).
Figure 2: Sequence coverage of a SARS-CoV-2 Omicron clinical sample with updated primer schemes
Integrative Genome Viewer visualization of read coverage across the SARS-CoV-2 genome (log scale). Amplicons were generated using NEBNext® VarSkip Short SARS-CoV-2 v2 primer pools, NEBNext VarSkip Short SARS-CoV-2 primer pools, NEBNext ARTIC SARS-CoV-2 primer pools (ARTICv3), MilliporeSigma ARTICv4 primer pools plus IDT ARTICv4.1 spike-in primers (ARTICv4.1), or MilliporeSigma ARTICv4 primer pools. The same Omicron variant SARS-CoV-2 viral gRNA clinical sample served as a template for all primer schemes except ARTICv3. Libraries were constructed using the NEBNext ARTIC SARS-CoV-2 FS Library Prep Kit (Illumina®) and sequenced on a MiSeq® instrument (2×75 bp). Coverage depth per base was determined, reads were down-sampled to 0.5M PE reads with seqtk and aligned to SARS-CoV-2 reference genome (NCBI, NC_045512) with Bowtie2.
Figure 3: Express and Standard workflows for the NEBNext ARTIC SARS-CoV-2 FS Library Prep Kit for Illumina with VarSkip Short v2 or NEBNext ARTIC primers
Where can I find guidelines and protocols for using the NEBNext ARTIC SARS-CoV-2 FS Library Prep Kit, including cDNA synthesis, cDNA amplification and library prep, and the Express protocol option with reduced cleanup steps?
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。
manualE7658 with VarSkip Short SARS-Co-V2 Primers v4point1
manualE7658 with VarSkip Short v2 SARS-CoV-2 Primers v5
工具 & 资源
Web 工具
NEBNext Selector
FAQs & 问题解决指南
FAQs
What is the difference between the NEBNext VarSkip Short v2 SARS-CoV-2 Primer Mixes and the original NEBNext VarSkip Short SARS-CoV-2 Primer Mixes?
What is the difference between the NEBNext VarSkip Short v2 SARS-CoV-2 Primer Mixes and the NEBNext ARTIC SARS-CoV-2 Primer Mixes?
Where can I find primer sequence information for NEBNext VarSkip Short v2 SARS-CoV-2 Primer Mixes and NEBNext ARTIC SARS-CoV-2 Primer Mixes?
Can the NEBNext ARTIC SARS-CoV-2 Primer Mixes and NEBNext VarSkip Short v2 SARS-CoV-2 Primer Mixes be added to the same targeted amplification reaction?
Which primer scheme should I use: NEBNext ARTIC SARS-CoV-2 Primer Mixes or NEBNext VarSkip Short v2 SARS-CoV-2 Primer Mixes?
Can the NEBNext VarSkip Short v2 SARS-CoV-2 Primers cover non-variant SARS-CoV-2 samples?
Which SARS-CoV-2 variants can the NEBNext VarSkip Short v2 SARS-CoV-2 Primers cover?
Can the NEBNext ARTIC Human Control Primer Pairs and NEBNext VarSkip Short v2 SARS-CoV-2 Primer Mixes be added to the same targeted amplification reaction?
How do I analyze VarSkip Short v2 SARS-CoV-2 sequencing data?
Are the sequences of the primers in the NEBNext ARTIC SARS-CoV-2 Primer Mixes the same as the ARTIC Network V3 primers?
What read length is recommended for sequencing of libraries generated using the NEBNext ARTIC SARS-CoV-2 FS Kit (Illumina)?
How many sequencing reads do I need for sequencing of libraries generated using the NEBNext ARTIC Kits for Illumina?
What Ct values are recommended for inputs used with the NEBNext ARTIC kits for Illumina?
How do I analyze data from libraries generated using the NEBNext ARTIC kits for Illumina?
Which NEBNext adaptors and primers are recommended for use with the ARTIC kits for Illumina?
Do I need to add PhiX to the sequencing run?
Do I need to normalize library concentration prior to sequencing?
What should the final loading concentration be for MiSeq® and NextSeq® 500/550 Illumina® sequencing platforms?
Does my total RNA sample need to be DNA-free prior to starting the ARTIC workflow?
