适用说明:本说明书适用于KAPA Stranded mRNA-Seq试剂盒 (07962193001和07962207001) 和KAPA mRNA Capture试剂盒 (07962231001和07962240001). 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
The KAPA Stranded mRNA-Seq Kit contains all of the buffers and enzymes required for poly(A) mRNA capture and construction of stranded mRNA-seq libraries from 100 ng – 4 μg of intact, total RNA via the following steps:
This kit provides all of the enzymes and buffers required for mRNA enrichment, cDNA synthesis, and library construction and amplification, but does not include RNA, adapters, or beads. KAPA Pure Beads and KAPA Adapters are sold separately. Reaction buffers are supplied in convenient formats comprising all of the required reaction components. This minimizes the risk of RNase contamination, ensures consistent and homogenous reaction composition, and improves uniformity among replicate samples. Similarly, a single enzyme mixture is provided for each step of the library construction process, reducing the number of pipetting steps. In order to maximize sequence coverage uniformity and to maintain relative transcript abundance, it is critical that library amplification bias be kept to a minimum. KAPA HiFi DNA Polymerase is designed for low-bias, highfidelity PCR, and is the polymerase of choice for NGS library amplification1,2,3,4. KAPA Stranded mRNA-Seq Kits include KAPA HiFi HotStart ReadyMix (2X) and Library Amplification Primer Mix (10X) for library amplification.
Product Applications
The KAPA Stranded mRNA-Seq Kit is designed for both manual and automated NGS library construction from 100 ng – 4 μg of intact, total RNA. The protocol is applicable to a wide range of RNA-seq applications, including: • gene expression • single nucleotide variation (SNV) discovery • splice junction and gene fusion identification • characterization of polyadenylated RNAs.
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.