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NxSeq® AmpFREE Low DNA Library Kit
NxSeq® AmpFREE Low DNA Library Kit
货号
14000-2
规格
48 rxn
存储温度
-20º C
产品详情
高效率，低输入，最低成本制备任意PCR片段文库试剂盒
- 低输入：对于限定的样品，至少需要输入75 ng剪切的DNA。
- 高效性：优化接头连接，使每个文库产生更多的测序片段，产生来自单一或多重文库的更高覆盖度和深度。
- 自由PCR: 防止引入PCR的偏好性，提供一致的覆盖度。
- 快速：2小时10分钟的操作流程，节省时间快速得到测序样品。
- 足以承担的：最优的价格和最好的材料。
您做新一代测序的原因是您需要大数据。为此，你需要把每一个样本都建成最好的文库来获得最多的信息量。那么为什么不选择一款好的DNA文库构建盒子来取代您所花费的时间，资源和昂贵的测序试剂。
The NxSeq® AmpFREE Low DNA Library Kit 可以让您建立最好的文库。我们优化了操作步骤的每一步确保在illumina测序仪上发挥最优的表现。此外盒子仅需要输入剪切DNA的量为75 ng。使用简化，容易的操作步骤在2小时的内构建出文库。
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比较好的数据
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- 高效文库
- 对于FFPE样本更大的读长
- 最优的测序数据
- GC偏好性最小化
- 低价，快速，易于操作
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Higher Efficiency Libraries
More Sequenceable DNA Fragments per Library = More Data
Figure 1. Percentage of library DNA with correctly ligated adaptors measured by qPCR. Duplicate libraries were prepped per kit/organism (Human, Staphylococcus aureus, Rhodobacter sphaeroides (1 library only), and E. coli) according to the manufacturer’s recommended input amounts and protocols. Adaptor ligation efficiency was measured by qPCR using the KAPA Library Quantification Kit (Complete ROX Low, cat #KK4873) and matching amplified library as an internal standard.
More Reads from Challenging FFPE Samples
Better Libraries Increase the Number of Reads per Library
| Library Kit | DNA Input | Total Number of Sequencing Reads Per Library | |
| Staphylococcus aureus | E. coli K12 | ||
| NxSeq® AmpFREE Low DNA Library Kit | 75 ng | 5,649,946 | 4,305,882 |
| Kapa Hyper Prep Kit | 250 ng | 4,838,726 (-15%) | 1,647,452 (-62%) |
| Illumina TruSeq DNA PCR-Free Library Prep Kit | 1 µg | 38,768 (-99%) | 1,543,558 (-64%) |
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Figure 2a. Number of sequencing reads generated per library after multiplexing and running on a MiSeq Instrument. DNA fragment libraries were prepped in parallel for each kit/organism according to the manufacturer’s recommended input amounts and protocols. Libraries were quantitated and normalized to 2 nM using the Bioanalyzer (size) and Qubit Fluorometer (amount). Equimolar amounts of each library were multiplexed and sequenced with a single MiSeq run using 2 ×150 bp chemistry. The number of sequencing reads obtained are shown as well as the percent reduction (%) in total reads compared to the appropriate NxSeq AmpFREE Kit results. |
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More Proof with Challenging FFPE Samples
| Library Kit | Sample Type | Input Amount | Total Reads |
Mapped Reads (repeat masked) |
| NxSeq® AmpFREE Low DNA Library Kit | Normal gDNA | 75 ng | 2,163,636 | 900,338 |
| FFPE DNA | 75 ng | 1,767,818 | 688,074 | |
| FFPE DNA | 150 ng | 1,706,714 | 656,658 | |
| Kapa Hyper Prep Kit | Normal gDNA | 250 ng | 1,567,276 (-28%) | 650,296 (-28%) |
| FFPE DNA | 250 ng | 1,270,870 (-28%) | 487,872 (-29%) | |
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Figure 2b. Number of sequencing reads generated from matching normal and FFPE gDNA sample libraries. DNA fragment libraries were prepped using the two indicated kits according to the manufacturer’s recommended input amounts and protocols. Libraries were constructed from normal gDNA (Biochain, Cat. No. D1234142-S02) and DNA extracted from a matching FFPE human kidney tissue (Biochain Cat. No. T2234142-S02) using the Qiagen AllPrep DNA/RNA FFPE Kit. The gDNA samples were sheared to ~250 bp before starting library construction. Final libraries were quantitated and normalized to 2 nM using the Bioanalyzer (size) and Qubit Fluorometer (amount). Equimolar amounts of each library were multiplexed and sequenced with a single MiSeq run using 2 × 150 bp chemistry. The number of sequencing reads obtained are shown as well as the percent reduction (%) in total and mapped reads compared to the corresponding NxSeq AmpFREE Kit results using 75 ng of input DNA. |
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Highly Mappable Reads (>92%) from Human, Staphylococcus andRhodobacter gDNA Sequencing
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Sequencing Stat |
Human | Staphylococcus |
Rhodobacter |
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Genome size, GC percentage |
~3 Gbp 45% GC |
2,821,361 33% GC |
4,602,977 69% GC |
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Raw reads |
3,131,114 |
1,260,836 |
3,900,174 |
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Mapped reads |
2,979,237 (95.15%) |
1,174,111 (93.12%) |
3,613,165 (92.64%) |
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Read length |
148.9 bp |
148.8 bp |
149.6 bp |
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Total bases |
443,767,447 |
174,694,261 |
540,403,552 |
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Genome fraction |
0.11 |
0.97 |
1.00 |
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Avg. coverage |
0.15X |
62X |
117X |
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Figure 3. Representative gDNA sequencing stats from three different organisms. Genomic DNA fragment libraries were generated using the NxSeq AmpFREE Low DNA Library Kit using 75 ng of sheared gDNA input from three organisms (human, Staphylococcus aureus, and Rhodobacter sphaeroides). The final libraries were quantitated and normalized to 2 nM final concentrations using the Bioanalyzer and Qubit fluorometer, and 5 µL of each library was run on a MiSeq using 2 x 150 bp chemistry and analyzed. |
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Minimal Bias Detected
Figure 4. Sequencing bias measured for three different organisms with varying GC content. DNA fragment libraries were generated from gDNA of three organisms with varying GC content (Staphylococcus aureus, 24% GC; E. coli K12, 50% GC; and Rhodobacter sphaeroides, 68% GC) according to the manufacturer’s recommended input amounts and protocols. Samples were quantitated using the Bioanalyzer and Qubit fluorometer and normalized to 2 nM final concentrations. Five µL of each library sample was sequenced on a MiSeq using 2 x 150 bp v2 chemistry and analyzed. Normalized coverage was calculated as the (average coverage of all windows with X% GC content) divided by the (overall average coverage).
Don’t spend 25-74% More per Library with Another Kit
Save Big with the NxSeq® AmpFREE Low DNA Library Kit
| PCR-Free Fragment Library Kit | Cat. No. |
Size (rxns) |
2016 US List Price |
Cost/Reaction |
Per Rxn Cost Increase vs. NxSeq® Kit |
|
|
Lucigen NxSeq® AmpFREE Low DNA Library Kit |
14000-1 | 12 | -- | -- | ||
| 14000-2 | 48 | -- | -- | 12 rxn | 48 rxn | |
| Kapa Hyper Prep Kit | KK8501 | 8 | -- | -- | +65% | +74% |
| KK8503 | 24 | -- | -- | +45% | +53% | |
| KK8505 | 96 | -- | -- | +25% | +32% | |
Faster Protocol with Significantly Less Hands-on Time
Save Time and Get Your Samples on the Sequencer Sooner!
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Ordering
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Manuals
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Resources
The NxSeq AmpFREE Low DNA Library Kit and Adaptors are only compatible with Illumina sequencers.
Each NxSeq AmpFREE Low DNA Library Kit contains Enzyme Mix (EM), 2X Buffer (2XB), Ligase (LIG) and Elution Buffer (EB). Adaptors must be purchased separately.
Each box of NxSeq Adaptors contains (12) different indexed Illumina-compatible adaptors with enough of each adapter for 4 library reactions. Box 1 contains adaptors 1-12 and Box 2 contains adapters 13-24.
The product manual is available under the Manuals tab.
新一代测序AmpFREE Low DNA 文库盒子和接头只与Illumina 测序仪匹配。
每一个新一代 AmpFREE Low DNA Library Kit包含 Enzyme Mix (EM), 2X Buffer (2XB), 连接酶 (LIG) and 稀释 Buffer (EB). 接头需要单独购买。
每个测序接头的盒子包括 12个不同的可与illumina兼容的接头，足够4个文库反应所需接头。盒子 1 含有接头1-12 盒子 2 contains 接头 13-24.
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