适用说明:本说明书适用于the KAPA HTP Library Preparation Kit (07961901001), 和 the KAPA HTP Library Preparation Kit for PCR-free workflows (07961910001). 友情提示: 1)本文档中的页码,附录均指原说明书中的页码及附录; 2)本文档包含原说明书中的产品描述,产品应用及实验操作流程、故障排查等主要信息,欲了解关于本产品详细说明及信息,建议参考原说明书 (点击面板上“下载PDF”可下载)。
Product Description
KAPA HTP Library Preparation Kit is designed for high- throughput library construction for Illumina sequencing, starting from fragmented, double-stranded DNA (dsDNA). The kit contains all of the enzymes and reaction buffers required for the following steps of library construction:
Efficient, cost-effective reaction cleanups and higher library yields are achieved through implementation of the “with-bead” strategy developed at The Broad Institute of MIT and Harvard and Foundation Medicine.1 The kit includes PEG/NaCl Solution for this purpose. Adapters and beads required for cleanups after end repair and library amplification are not included. However, KAPA Pure Beads and KAPA Adapters are sold separately. In order to maximize sequence coverage uniformity, it is critical to minimize library amplification bias. KAPA HiFi DNA Polymerase is designed for low-bias, high- fidelity PCR, and is the reagent of choice for NGS library amplification.2,3,4,5 KAPA HTP Library Preparation Kits include KAPA HiFi HotStart ReadyMix (2X), a ready-to- use PCR mix comprising all the components for library amplification—except primers and template. Kits also include Library Amplification Primer Mix (10X), designed for the high-efficiency amplification of Illumina libraries flanked by adapters containing the P5 and P7 flow cell sequences. A kit without the amplification module (KK8235) is available for PCR-free workflows. They may also be combined with KAPA HiFi Real-time Library Amplification Kits, or with KAPA HiFi HotStart Uracil+ ReadyMix Kits for the amplification of bisulfite-converted libraries.
Product Applications
KAPA HTP Library Preparation Kits are ideally suited for high-throughput NGS library construction workflows that require end repair, A-tailing, adapter ligation and library amplification (optional). The kit is designed for library construction from a wide range of sample types and inputs (10 ng – 5 µg). For small genomes, cell-free/circulating tumor DNA and lower complexity samples such as ChIP DNA, amplicons or cDNA (for RNA-seq), successful library construction has been achieved from lower inputs (~100 pg or more). The protocol is automation friendly and may be incorporated into workflows for a wide range of NGS applications, including: ● whole-genome, shotgun sequencing ● whole exome or targeted sequencing, using Roche SeqCap EZ, Agilent SureSelect, Illumina TruSeq, IDT xGen Lockdown Probes, or other hybridization capture systems ● ChIP-seq ● RNA-seq (starting with cDNA) ● methyl-seq (in combination with the KAPA HiFi HotStart Uracil+ ReadyMix for library amplification). Specific guidelines for the construction of libraries for target capture using the Roche SeqCap EZ system may be found in the Appendix (p 17).
Process Workflow
Library Construction Protocol
If you have any questions/comments about this protocol, you are highly recommended to post here. We will invite the authors of this protocol as well as some of its users to address your questions/comments. To make it easier for them to help you, you are encouraged to post your data including images for the troubleshooting.