SureSeq™ NGS Library Preparation Kit
The complete library preparation solution for unparalleled next generation sequencing (NGS) results.
The SureSeq NGS Library Preparation Kit generates NGS libraries suitable for the capture of targeted genomic regions using hybridisation. With a streamlined workflow, significantly reduced hands-on steps and hybridisation times as low as 30 minutes, SureSeq offers all of the benefits of hybridisation in as little as a 1-day workflow. For increased convenience and flexibility, the SureSeq workflow can be performed with an enzymatic or Covaris® DNA fragmentation and Speedvac® or bead concentration steps, whilst still delivering libraries of the highest quality. The inclusion of the SureSeq Hyb & Wash buffer, optimised for use with SureSeq NGS panels, simplifies this key step while offering excellent coverage uniformity and reproducibility. Additionally, a new automation workflow enables standardisation of OGT’s library preparation, hybridisation and washing steps for up to 96 samples in a single batch, improving laboratory productivity.
The SureSeq Library Preparation Kit offers:
- Greater trust in your data - high performance with low duplication rates, high sequence quality and high percentage of on-target bases
- Simpler hybridisation - all components of the SureSeq Hyb & Wash buffer are ready-to-use with no requirement for multiple wash buffers
- Rapid process - streamlined protocol, minimal manual handling, automation and a rapid hybridisation step offers increased reliability as well as throughput
- Reliable results - NGS targeted panel, complete NGS library preparation solution and powerful NGS analysis software are fully optimised for excellent results
Greater trust in your data
NGS data is increasingly being relied upon as a front-line technology for the generation of data for scientific and medical research. A combination of quality metrics are used to give confidence that variants called are real and the number of false positive calls are low. The SureSeq NGS Library Preparation Kit delivers high performance in the quality metrics that really matter, giving more reliable, more trustworthy data.
Low levels of duplication
High duplication rates reduce the complexity of your NGS library and can lead to poor coverage of targeted genomic regions even though average coverage rates appear high.
The SureSeq NGS Library Preparation Kit gives exceptionally low levels of duplication (Figure 1). This ensures more accurate calling, more even coverage and higher levels of confidence in the data produced.
Figure 1. The SureSeq NGS Library Preparation Kit delivers low levels of sequence duplication. The duplication rates are shown for samples fragmented by mechanical or enzymatic methods. Samples were prepared using the SureSeq NGS Library Preparation kit and hybridised with a SureSeq myPanel Custom Myeloid 49 Gene Plus panel.
High quality sequence data
For complete confidence in your data, the sequence quality scores should be high and consistent across the length of each read. The SureSeq NGS Library Preparation Kit delivers exceptional sequence quality scores even 150 bases into the read (Figure 2). This reduces the requirement for post-acquisition bioinformatics, such as read size reduction, to remove poor quality sequence.
Figure 2. The SureSeq NGS Library Preparation Kit delivers high sequence quality. Data shown was generated using 4 samples of human genome DNA, the OGT SureSeq Myeloid Panel, and the SureSeq NGS Library Preparation Kit.
High levels of on-target bases
In order to maximize the amount of useful information that can be obtained for a given sequencing lane, high coverage depth and low coefficients of variation are desirable. The benefits of superior levels of specific sequence capture include increased sequence complexity and greater depth of coverage of the target regions. The SureSeq NGS Library Preparation Kit delivers excellent reproducibility (Figure 3).
Figure 3. High depth of coverage and a low coefficient of coverage variation from 16 replicates. A SureSeq myPanel Myeloid 49 gene plus and SureSeq Library Preparation Kit were utilised in the evaluation of 336 coding targets in the custom panel (excluded targets on X or Y chromosome) to illustrate the low CV (coefficient of variation) and excellent uniformity of SureSeq.
Simpler hybridisation with the SureSeq NGS Hyb & Wash Kit
The SureSeq NGS Hyb & Wash kit is an easy to use, robust and high performing hybridisation solution, containing all the components required to perform the hybridisation and wash steps in SureSeq sequence capture protocols. All components in the kit are ready-to-use, eliminating the requirement to dilute buffers and the possibility of cross-contamination during buffer preparation. There is no need to use multiple wash buffers, as in other manufacturer’s kits, the simplified protocol utilises just a single wash buffer. With hybridisation times as little as 30 minutes, it offers excellent quality sequencing data, when used in conjunction with the SureSeq NGS Library Preparation Kit and SureSeq NGS panels (Figure 4).
Figure 4. Superior performance. Comparison of the mean target coverage generated using the SureSeq Hyb & Wash Buffer Kit (light blue) compared to an alternative supplier kit (dark blue). [A] SureSeq myPanel custom CLL panel and [B] SureSeq myPanel custom CLL plus CNV panel.
Rapid process with streamlined protocol
By extensive testing of enzymes and buffer optimisation, it has been possible to streamline the standard library preparation protocol to minimise the number of hands-on steps and the overall processing time (Figure 5). For added convenience, OGT now offers Dynabeads™ M270 Streptavidin and AMPure® XP beads for use with SureSeq NGS panels. While the addition of a workflow for automation on the Agilent Bravo A® Automated Liquid Handling Platform offers greater flexibility and productivity for your laboratory.
- End repair and ligation are combined into a single step
- Purification steps, where material is lost, are reduced
- Fewer steps reduce handling errors for increased reliability
- Reduced technician time and cost and increased sample throughput
- Workflow amenable to automation
Figure 5. A streamlined protocol, including enrichment by hybridisation. With hybridisation times as low as 30 minutes, the procedure can be completed in as little as one day with minimal handling time.
Reliable results with SureSeq targeted NGS panels and analysis software
The SureSeq NGS Library Preparation Kit and Hyb & Wash Kit work hand-in-hand with SureSeq targeted enrichment panels, ensuring you get the most sensitive and reproducible variant detection, (Figure 6), and industry-leading coverage uniformity.
SureSeq Interpret is a powerful and easy-to-use next generation sequencing analysis solution. Sequencing data can be quickly processed to deliver accurate identification of single nucleotide variants (SNVs) and Indels as well as structural variants such as copy number variants (CNVs) and internal tandem duplications (ITDs). Coupled with a comprehensive and powerful filtering framework, the software delivers accurate mutation calling with 100% sensitivity and 99.9% specificity at >1% variant allele frequency (VAF)*. It is designed to work with Oxford Gene Technology’s (OGT) extensive range of SureSeq NGS panels and offers flexible accessibility to analyse your data whether through a stand-alone computer†, laboratory server or another web enabled device.
Figure 6. Excellent reproducibility with low allele frequency detection and CV distribution. The SureSeq workflow and software were validated using OncoSpan Reference Standards (Horizon Discovery). A SureSeq myPanel NGS Custom panel was utilised to detect 46 variants in 15 genes present in the OncoSpan Reference Standard. Three different DNA inputs with three independent replicates were processed in parallel. Data generated was analysed using SureSeq Interpret Software.
|SureSeq NGS Library Preparation Complete Solution (16)
||Bundle of 1x SureSeq library preparation kit (16), containing adaptors, PCR primers and enzymes, 1x SureSeq NGS Index Kit – Collection A, 1x SureSeq Hyb & Wash Kit (16), 1x Dynabeads M270 Streptavidin (2ml) and 1x AMPure XP beads (10ml). Sufficient for 16 samples
||Get a quote|
|SureSeq NGS Library Preparation Complete Solution (48)
||Bundle of 3x SureSeq NGS Library Preparation Kit (16), containing adaptors, PCR primers and enzymes, 1x SureSeq NGS Index Kit – Collection B, 3x SureSeq NGS Hyb & Wash Kit (16), 3x Dynabeads M270 Streptavidin (2ml) and 3x AMPure XP beads (10ml). Sufficient for 48 samples
||Get a quote|
|SureSeq NGS Library Preparation and Hyb & Wash Kit (16)
||Bundle of 1x SureSeq NGS Library Preparation Kit (16), containing adaptors, PCR primers and enzymes, 1x SureSeq NGS Index Kit – Collection A and 1x SureSeq Hyb & Wash Kit (16). Sufficient for 16 samples
||Get a quote|
|SureSeq NGS Library Preparation and Hyb & Wash Kit (48)
||Bundle of 3x SureSeq NGS Library Preparation Kit (16), containing adaptors, PCR primers and enzymes, 1x SureSeq NGS Index Kit – Collection B and 3x SureSeq Hyb & Wash kit (16). Sufficient for 48 samples
||Get a quote|
|SureSeq NGS Library Preparation Kit (16)
||Bundle of 1 x library preparation kit (16), containing adaptors, PCR primers and enzymes sufficient for 16 samples and 1 x SureSeq NGS Index Kit – Collection A
||Get a quote|
|SureSeq NGS Library Preparation Kit (48)
||Bundle of 3 x library preparation kit (16), containing adaptors, PCR primers and enzymes sufficient for 48 samples and 1 x SureSeq NGS Index Kit – Collection B
||Get a quote|
|SureSeq NGS Hyb & Wash Kit (16)
||Hybridisation buffer, Wash buffer, Cot and blocking oligos. Sufficient for 16 samples
||Get a quote|
|SureSeq NGS Hyb & Wash Kit (48)
||Bundle of 3x SureSeq NGS Hyb & Wash Kit (16), containing Hybridisation buffer, Wash buffer, Cot and blocking oligos. Sufficient for 48 samples
||Get a quote|
|Sure Seq FFPE DNA Repair Mix (16 reactions)||Enzyme mix and buffer sufficient for 16 FFPE DNA samples||500079‡||Get a quote|
|Dynabeads M270 Streptavidin, 2ml||Sample capture beads, sufficient for 20 samples||500080**||Get a quote|
|AMPure XP beads, 10ml||Sample purification beads, sufficient for 16 samples||500081**||Get a quote|
* Sensitivity and specificity determined using Horizon Discovery OncoSpan and TruQ7 and HapMap (NA12878) standards.
† Computer specification: RAM: min 16Gb, suggested 24Gb; HDD: min 500Gb, suggested 2TB; CPU: 8+ logical cores at 2+ GHz; OS: (Windows 7 or newer), Virtualization (VT-x) needs to be enabled in the BIOS.
‡ The SureSeq™ FFPE DNA Repair Mix can only be purchased in conjunction with SureSeq NGS panels, not as a standalone product.
**Only for use with SureSeq NGS panels
SureSeq: For Research Use Only; Not for Diagnostic Procedures. This webpage and its contents are © Oxford Gene Technology IP Limited – 2019. All rights reserved. OGT™ and SureSeq™ are trademarks of Oxford Gene Technology IP Limited. The SureSeq NGS Library Preparation Kit was jointly developed between Oxford Gene Technology and Bioline Reagents Limited. SpeedVac® is a registered trademark of Thermo Fisher Scientific, Covaris™ is a registered trademark of Covaris, Inc., Bravo® is a registered trademark of Agilent Technologies, Inc., Dynabeads is a trademark of Thermo Fisher Scientific and AMPure® is a registered trademark of Beckman Coulter Inc.
Evaluation of enzymatic DNA digestion as an alternative to mechanical DNA fragmentation (sonication) for targeted NGS using the SureSeq™ Myeloid Panel
DNA fragmentation is a crucial first step in the preparation of libraries for NGS. In this application note, Oxford Gene Technology has evaluated an alternative method of fragmentation using the NEBNext® dsDNA Fragmentase®.
Improving experimental reproducibility through automated hybridisation-based NGS library preparation
In this application note, an Agilent Bravo A Automated Liquid Handling Platform was configured to run the SureSeq NGS library preparation protocol. The results demonstrate marked improvement not only in hands-on-time, but also a number of quality metrics
Selecting the best NGS enrichment assay for your needs
With NGS now in routine use for a broad range of research and clinical applications, this application note details the value of making the correct choice for the initial sequence enrichment step.
Optimised, 1-day hybridisation-based NGS protocol yields 1% variant detection in MPN samples, as quickly and cost-effectively as multiplex PCR
Presented at AMP 2016, this poster outlines how the SureSeq™ Core MPN Panel can accurately detect alleles down to 1% variant allele fraction (VAF) in JAK2 (V617F) at a read depth of >1000x, facilitating reliable detection.
The accurate detection by next-generation sequencing (NGS) of difficult to sequence genes (CALR, CEBPA, FLT3) associated with myeloid disorders using a hybridisation-based enrichment approach
Presented at CGC 2017, this poster highlights the excellent uniformity of coverage obtained from the hybridisation-based enrichment using the SureSeq myPanel NGS Custom AML Panel.
The analysis of myeloproliferative neoplasm samples using a rapid (30 minute) hybridisation-based enrichment protocol for next-generation sequencing (NGS)
Presented at the CGC 2017 annual summer meeting in Denver, USA, this poster illustrates the excellent quality data generated by the OGT 1-day hybridisation-based SureSeq LPK protocol in combination with the SureSeq Core MPN Panel.
The application of a hybridisation-based next-generation sequencing (NGS) enrichment panel for the analysis of key genes involved in ovarian and breast tumours using DNA from FFPE samples
The application of a hybridisation-based NGS enrichment panel for the analysis of somatic variants in tumour samples and reference standards
Presented at AGT 2017, this poster outlines the application of a hybridisation-based NGS enrichment panel for the analysis of solid tumour somatic variants, demonstrating 100% concordance in variant detection in both genomic and formalin-compromised DNA.
The application of a one-day hybridisation-based enrichment protocol for NGS incorporating a rapid (30 minute) hybridisation step
Presented at AGT 2017, this poster outlines how OGT has optimised a one-day hybridisation-based enrichment protocol for NGS incorporating a rapid 30 hybridisation step.
The use of a hybridisation-based NGS enrichment panel for the confident identification of a broad range of low frequency variants from as little as 50ng of challenging clinical research FFPE samples
Presented at AMP 2016, this poster outlines how the SureSeq FFPE DNA Repair Mix significantly improves NGS library yields, with an increase of mean target coverage (increased by >2.2 fold), resulting in more meaningful data.
SureSeq myPanel™ NGS Custom Cancer Panels Full Gene List
We now have 120 genes available for our SureSeq myPanel NGS Custom Cancer Panels. View and download a complete list of available cancer gene content.
The role of NGS in stratified cancer medicine
In this white paper, two Clinical Scientists, Dr Matthew Smith and Dr George Burghel, share their views on the use of NGS in cancer genomics and its integration into the laboratory.
Understanding myeloid disorders with next-generation sequencing
How OGT’s SureSeq™ Myeloid Panel helps researchers identify and decipher the complex genetic origins of myeloproliferative disorders
SureSeq myPanel™ NGS Custom Breast Cancer panel
SureSeq myPanel™ NGS Custom Cancer Panels
SureSeq myPanel™ NGS Custom Colorectal Cancer Panel
SureSeq myPanel™ NGS Custom Melanoma Cancer Panel
SureSeq™ FFPE DNA Repair Mix
SureSeq™ NGS Library Preparation Kit
SureSeq™ Ovarian Cancer Panel