Qiagen/QIAseq Methyl Library Kit/Enzymes, buffers, adapters (up to 24 sample indexes) and QIAseq Beads for construction of 24 libraries from bisulfite-treated DNA/180502
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商品介绍
QIAseq Methyl Library Kit
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For robust library preparation for DNA methylation analysis
- High library yield and mapping rates
- Low bias and error rates
- Wide range of DNA input
- Compatible with cfDNA samples
- Suitable for enriched fragmented samples (e.g., RRBS, MeDIP)
The QIAseq Methyl Library Kit is a library construction kit usedfor preparing libraries from bisulfite-treated DNA samples for whole genome methylation studies using Illumina platforms. It is developed for researchers who need a robust solution to interrogate methylation events in their biological samples. The kit combines several library preparationreactions into one streamlined stepthat enables completelibrary preparation in 2.5 hours and allows thefull workflow, including bisulfite conversion, to be performed inless thana day. Unlike other kits that requireconsiderable DNA input, the QIAseq Methyl Library kitallows DNA inputs as low as 100 pg to deliver high-quality, high-yield libraries for use withIllumina platforms.The kit includes all requiredcomponents to go from bisulfite-treated DNA to libraries: enzymes, buffers and reagents for double-strand synthesis, end-polishing of bisulfite-converted DNA, ligation of Illumina adapters, library purification with QIAseq Beads and library amplification with the VeraSeq ULtra DNA Polymerase.
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Cat No./ID:180502 QIAseq Methyl Library Kit (24) $1,253.00 Enzymes, buffers, adapters (up to 24 sample indexes) and QIAseq Beads forconstruction of 24 librariesfrom bisulfite-treated DNA |
Product Details
Performance
High yield
The superior chemistryof the QIAseq Methyl Library Kit enables high library yields,usingas littleas 100 pg of input DNA for library construction,from alownumber of amplification cycles (Figure High library yield and mapping rate).High mapping rates
One of the major challenges in the use of NGS for methylation analysis is low mapping rates, resulting in inefficient use of the sequencing run. The QIAseq Methyl Library Kit overcomes this challenge by delivering mapping rates greater than 90%, with an optimal DNA input range of 1–100 ng (Figure High library yield and mapping rate).Interrogates CpG methylation with low bias
Accurate methylation analysis is mostly affected by the input material and unbiased conversion of the bisulfite-treated and heavily-fragmented DNA into library molecules. The high degree of bisulfite conversion provided by EpiTect Fast and efficient repair of the bisulfite-treated DNA, coupled with efficient ligation of adapters, make accurate methylation analysis possible, even from very low DNA input (Figure Accurate methylation state calling).Low error rates
With an optimal input range of 1–100 ng DNA, the QIAseq Methyl Library Kit, which includes the high-fidelity VeraSeq ULtra DNA Polymerase, reduces error rates for more accurate assessment of methylation events.Principle
The QIAseq Methyl Library Kit contains enzyme mixes and buffers forpreparing bisulfite-treated single-stranded DNA for ligation of adapters that allow sequencing on Illumina platforms and amplification of the libraries. Magnetic beads are includedfor library purification. In a simplified workflow, the kit uses an enzyme mix and random short primers for double strand synthesis of single-stranded bisulfite-treated DNA. In the same reaction setup, enzymes willend-polish and prepare the ends of the fragments for subsequent adapter ligation.The reactions, double-strand synthesis, DNA end-polishing and adapter ligation, take place in the same tube, thereby minimizing sample loss. The ligation step is followed by a purification step using theincluded QIAseq Beads. Library amplification is performed using thehigh-fidelity VeraSeq ULtra DNA Polymerase, which isan engineered, ultra-thermostable, uracil-literate polymerase designed to maximize the speed, accuracy and length of DNA synthesis during sequencing template preparation. The result is a novel enzyme that can read through uracil, extend a kilobase of sequence in 15 seconds and has an accuracy 25 times higher than Taq DNA Polymerase. This enables the amplification of WGBS libraries without sequence bias.Dual-bar-coded, plate-format adapters are included in QIAseq Methyl DNA Library Kits. Each adapter well contains a single-use adapter consisting of a unique combination of two eight-nucleotide identification bar codes. By combining one of eight D5 bar codes and one of three D7 bar codes in each ready-to-use adapter, this kit supports up to 24-plexing prior to sequencing (Figure The QIAseq Methyl Library Kit workflow).
Procedure
Before library construction, DNA samples undergo bisulfite treatment with dedicated EpiTect Fast protocols for the complete conversion of non-methylated cytosines. Library construction begins with double-strand synthesis and end-polishing followed by adapter ligation. Following library construction, the reaction cleanup and removal of adapter-dimersis achieved using QIASeq Beads. After library amplification, a final cleanup step is performed with QIASeq Beads. After sequencing on anIllumina platform, bioinformatic analysis issimplified byusing the Bisulfite Sequencing plugin (version 2.0 or later) for Biomedical Genomics Workbench and CLC Genomics Workbench for non-directional libraries (Figure Sample to Insight workflow for the analysis of methylation events).
Applications
The QIAseq Methyl Library Kit produces high-quality libraries for the analysis of CpG methylation events from a wide range of samples, including gDNA, cfDNA and FFPE, as well as fragmented and enriched CpG fragments.Bisulfite conversion leads to DNA damage and makes bisulfite sequencing of heavily fragmented samples very difficult.The robust chemistry of the EpiTect Fast Kits and optimized protocols of the QIAseq Methyl Library Kit allow sequencing of difficult samples such as ccfDNA (Figure High percentage of mapped reads and accurate methylation calls with low-input circulating cell-free DNA).
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品牌介绍
QIAGEN是一家专业化致力于生物分子样品制备解决方案的跨国经营企业，总部位于德国。1984年，QIAGEN在德国成立，1996年在美国纽约纳斯达克上市。QIAGEN拥有超过1000项专利和认可证明，在18个国家设立了分公司，代理商服务国超过40个，在全球有超过400000的用户。QIAGEN提供的产品超过500类，包括各种试剂，耗材和自动化纯化工作站。这些产品用于样品采集，稳定，核酸或蛋白的分离，纯化和检测中，不仅广泛的应用于科研领域的各个方面，在生物技术，制药，法医研究，食品安全检测，畜牧业和分子诊断领域也得到了广泛的应用。QIAGEN产品的卓越品质和在应用中的出色表现使得其成为样品处理中标准的代名词。
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