·产品描述

       DNA文库的制备是下一代测序的必要步骤.为了产生精确的DNA测序信息，需要制备高质量和足量的DNA文库.同时，下一代测序技术在持续进步，DNA文库的制备也相应优化了.大多数现行的方法耗时、昂贵和不便.一些方法通过连接dA尾或者甚至把连接一步完成加快了实验速度，但是有很多G尾形成或是连接时形成很多DNA串联物，或者有很高的插入偏差.这些副作用设置导致了DNA文库效率低下和不准确.一个理想的DNA文库的制备方法应当是在速度、便捷、适合小样本、经济和准确之间取得平衡.为了解决这个问题，Epigentek开发了EpiNext DNA文库制备试剂盒.

       The EpiNext DNA文库制备试剂盒（Illumina） 是一组完整的、被优化的试剂组合，可以进行DNA文库的制备.制得的DNA文库可以通过采用一个 Illumina 序列进行下一代测序技术测序，包括基因组DNA测序、染色质共沉淀测序、甲基化DNA免疫共沉淀/羟甲基化DNA免疫共沉淀芯片测序、重亚硫酸盐测序和基因组目标区域重测序.优化的方案和组分可以进行非条形码序列（单个）和条形码序列（多重）DNA文库的快速、小偏差构建成为可能。

·产品特点

1.快速的试验流程.从片段化的DNA到片段大小选择只要2.5h. 

2. 非常便捷.试剂盒含有制备DNA文库步骤所必须的必要组分 

3.误差小.超级HiFi扩增和选择性的无PCR步骤使得实验者可以在最小误差的情况下得到最大的DNA文库产率 

4.灵活性.优化的方案和组分可以进行非条形码序列（单个）和条形码序列（多重）DNA文库的快速、小偏差构建成为可能

·产品说明书

The EpiNext™ DNA Library Preparation Kit  (Illumina) is a complete set of optimized reagents to carry out a successful DNA library preparation. The kit is suitable for preparing a DNA library for next generation sequencing applications using an Illumina sequencer, which includes genomic DNA-seq, ChIP-seq, MeDIP/hMeDIP-seq, bisulfite-seq, and targeted re-sequencing. The optimized protocol and components of the kit allow both non-barcoded (singleplexed) and barcoded (multiplexed) DNA libraries to be constructed quickly with reduced bias. The kit has the following advantages:

• Fast and streamlined procedure - The procedure from fragmented DNA to size ion is less than 2 h 30 min.  Only one clean-up between each step, thereby saving time and preventing handling errors, as well as loss of valuable samples. Gel-free size ion further reduces the preparation time.
• Highly Convenient - The kit contains all required components for each step of DNA library preparation, which are sufficient for end repair, dA tailing, ligation, clean-up, size ion and library amplification, thereby allowing the library preparation to be streamlined with the most reliable and consistent results. 
• Minimized bias - Ultra HiFi amplification and an optional PCR-free step enable the user to achieve reproducibly high yields of DNA library with minimal sequence bias and low error rates.
• Flexibility - Can be used for both non-barcoded (singleplexed) and barcoded (multiplexed) DNA library preparation. Uses various dsDNA including fragmented dsDNA isolated from various tissue or cell samples, dsDNA enriched from ChIP reactions, MeDIP/hMeDIP reactions, or exon capture. Broad range of input DNA from 10 ng to 1 µg. PCR-free library preparation can be performed with use of 500 ng or more input DNA.

Background Information
DNA library preparation is a critical step for next generation sequencing (NGS). To generate accurate sequencing data for NGS, the prepared library DNA should be sufficient in yield and of high quality. Also, as NGS technology is continuously improving, DNA library preparation is required to be optimized accordingly.  Most of the currently used methods are unfortunately time-consuming, expensive, and inconvenient. Some of the methods are relatively quick by combining end repair and dA tailing or even ligation in one-step, but have been shown to generate significant G tailing or form concatmers at the ligation step or have high insertion bias. These side reactions eventually result in the prepared DNA library being less efficient and inaccurate. An ideal DNA library preparation method should be balanced in speed, convenience, small sample-suitability, cost-effectiveness, and accuracy. To address this issue, Epigentek offers the EpiNext DNA Library Preparation Kit.

Principle & Procedure
This kit incluldes all reagents required at each step to carry out a successful DNA library preparation. In the library preparation, DNA is first fragmented to the appropriate size (about 300 bp peak size). The end repair of the DNA fragments is performed and an A-overhang is added at the 3'-end of each strand.  Adaptors are then ligated to both ends of the end repaired/dA tailed DNA fragments for amplification and sequencing. Fragments are then size ed and purified with MQ beads, which allows for quick and precise size ion of DNA. Size-ed DNA fragments are then amplified with a high-fidelity PCR Mix which ensures maximum yields from minimum amounts of starting material and provides highly accurate amplification of library DNA with low error rates and minimum bias. 

Starting Materials
Starting materials can include fragmented dsDNA isolated from various tissue or cell samples, dsDNA enriched from ChIP reactions, MeDIP/hMeDIP reaction, or exon capture. DNA should be relatively free of RNA since large fractions of RNA will impair end repair and dA tailing, resulting in reduced ligation capabilities. Input amount of DNA can be from 5 ng to 1 µg. For optimal preparation, the input amount should be 100 ng to 200 ng. For amplification-free, 500 ng or more is needed.