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Product Description
The KAPA Stranded RNA-Seq Kit with RiboErase (HMR) contains all of the buffers and enzymes required for depletion of ribosomal RNA (rRNA) followed by construction of stranded RNA-seq libraries from 100 ng – 1 μg of total RNA via the following steps:
The kit provides all of the enzymes and buffers required for rRNA depletion, 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, high-fidelity PCR, and is the polymerase of choice for NGS library amplification1,2,3,4. The KAPA Stranded RNA-Seq Kit with RiboErase (HMR) includes KAPA HiFi HotStart ReadyMix (2X) and Library Amplification Primer Mix (10X) for library amplification.
Product Applications
The KAPA Stranded RNA-Seq Kit with RiboErase (HMR) is designed for both manual and automated NGS library construction from 100 ng – 1 μg of total RNA. The kit depletes both cytoplasmic (5S, 5.8S, 18S, and 28S), and mitochondrial (12S and 16S) rRNA species. The protocol is applicable to a wide range of RNA-seq applications, including: • gene expression analysis of high- and low-quality RNA samples (e.g., extracted from FFPE tissue) • single nucleotide variation (SNV) discovery • splice junction and gene fusion identification • characterization of non-polyadenylated RNAs, including non-coding and immature 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.