Product Overview
RNA sequencing projects often need both conventional libraries and strand-specific libraries, and buying a separate kit for each adds cost and complexity. The Hyaseq Dual-mode RNA Library Prep Kit covers both in a single kit, and works on both the Illumina and MGI sequencing platforms. The kit is a total RNA library preparation kit that includes RNA fragmentation reagents, reverse transcription reagents, conventional and strand-specific ds-cDNA synthesis reagents, and library amplification reagents. Sequencing libraries can be constructed after an mRNA purification kit or an rRNA removal kit, so the upstream enrichment step can be chosen to match the sample and the sequencing goal. The “dual-mode” design is in the two-strand synthesis module, which comes with two buffers: one for routine (non-strand-specific) libraries, and one for strand-specific libraries. In the strand-specific buffer, dTTP is replaced with dUTP, so dUTP is incorporated into the second strand of cDNA. The high-fidelity DNA polymerase in the kit cannot amplify DNA templates that contain uracil, so the second strand is not amplified and strand specificity is achieved. Platform compatibility is handled with two primer mixes, one for Illumina and one for MGI, so the same kit supports both. The workflow is also simplified because two-strand synthesis, end repair, and A addition are combined into one step. All reagents provided have undergone strict quality control and functional verification, ensuring the stability and reproducibility of library preparation to the greatest extent.
Product Features
- Conventional and strand-specific libraries in one kit: Two two-strand synthesis buffers let you choose dNTP-based synthesis for non-strand-specific libraries, or dUTP-based synthesis for strand-specific libraries, without buying a separate kit for each mode.
- Illumina and MGI compatibility: A primer mix for Illumina and a primer mix for MGI are provided, so one kit supports library preparation for both platforms.
- Simplified workflow with uniform transcript coverage: Two-strand synthesis, end repair, and A addition are combined into one step, and the resulting sequencing data has excellent quality with more uniform coverage of transcript regions.
FAQ
Q1: How does the kit produce strand-specific libraries?
The strand-specific two-strand synthesis buffer replaces dTTP with dUTP, so dUTP is incorporated into the second strand of cDNA. The high-fidelity DNA polymerase used for library amplification cannot amplify templates that contain uracil, so the second strand is not amplified, and the library keeps the strand information of the original RNA.
Q2: Can I use the same kit for both conventional and strand-specific libraries?
Yes. The two-strand synthesis module comes with two buffers, one for routine libraries and one for strand-specific libraries. You choose the buffer that matches your project, and the rest of the workflow is the same.
Q3: Which sequencing platforms and upstream RNA preparation methods is the kit compatible with?
The kit is designed for the Illumina and MGI sequencing platforms, using a platform-specific primer mix for each. Libraries can be constructed after mRNA purification or rRNA removal, so the upstream enrichment step can be chosen to match your sample type.
