Hieff NGS™ Ultima Dual-mode mRNA Library Prep Kit is a mRNA transcriptome library construction kit compatible with Illumina and MGI high-throughput sequencing platforms. Compared with the traditional library construction method, this product combines cDNA second strand synthesis with Endprep and dA-tailing, which greatly reduces the time for library construction and simplifies the operation. The two-strand synthesis module is equipped with two buffers to meet the need for a conventional library or strand-specific library. This product is compatible with 10 ng-4 μg of input total RNA from eukaryotes. After mRNA isolation, fragmentation, double-stranded cDNA synthesis, end repair, dA-tailing, adapter ligation, and library amplification, the total RNA sample is finally converted into a library suitable for sequencing on the Illumina or MGI platform.
Hieff NGS™ Ultima Dual-mode mRNA Library Prep Kit contains two independent modules. The core components of BOX-I are oligo (dT) magnetic beads required for mRNA purification. BOX-II contains mRNA fragmentation reagents, reverse transcription reagents, conventional and chain-specific ds-cDNA synthesis reagents, and others required for subsequent library construction. dTTP is replaced with dUTP in the strand-specific two-strand synthesis Buffer so that dUTP is incorporated into the second strand of cDNA. The high-fidelity DNA polymerase used in this kit cannot amplify the DNA template containing uracil, which achieves strand specificity. All reagents provided have undergone strict quality control and functional verification, ensuring the stability and reproducibility of library construction to the greatest extent.
- One kit, four types of libraries: Illumina platform strand specific library; Illumina platform regular RNA library; MGI platform strand specific library; Illumina platform regular RNA library;
- Streamline the process: end repair and dA-tailing are combined in one step, simplifying the operation steps
- Wide scope of application: applicable to samples containing polyA tails of RNA from different sources, total RNA 10 ng-4 μ g
- High gene detection rate
- Stable quality: strict batch performance and stability quality control
- Gene expression detection: detect the mRNA expression in the sample
- Detection of SNP, Indel, etc
- Identification of fusion gene
- Analysis of alternative splicing events
|Library type||mRNA library|
|mRNA isolation method||Oligo dT Beads|
|Total RNA input amount||10 ng-4 μg|
|Library preparation total time (hr)||~5h|
|Instrument compatibility||Illumina or MGI Platforms|
|Recommended application||Gene expression|
|BOX-I||12603-A||mRNA Capture Beads||1.2 mL||4.8 mL|
|12603-B||Beads Binding Buffer||1.2 mL||4.8 mL|
|12603-C||Beads Wash Buffer||15 mL||60 mL|
|12603-D||Tris Buffer||1.2 mL||4.8 mL|
|BOX-II||12309-A||Frag/Prime Buffer||450 μL||2×900 μL|
|12309-B||1st Strand Enzyme Mix||48 μL||192 μL|
|12309-C||Strand Specificity Reagent||150 μL||580 μL|
|12309-D||2nd Strand Buffer (dNTP)||720 μL||2×1440 μL|
|12309-E||2nd Strand Buffer (dUTP)||720 μL||2×1440 μL|
|12309-F||2nd Strand Enzyme Master Mix||120 μL||480 μL|
|12309-G||Ligation Library length distribution detection||720 μL||2×1440 μL|
|12309-H||Novel T4 DNA Ligase||120 μL||480 μL|
|12309-I||2×Super Canace™ II High-Fidelity Mix||600 μL||2×1200 μL|
Shipping and Storage
The Hieff NGS™ Ultima Dual-mode mRNA Library Prep Kit components in Box I are shipped with ice packs and can be stored at 2℃~8℃ for one year.
The Hieff NGS™ Ultima Dual-mode mRNA Library Prep Kit components in Box II are shipped with dry ice and can be stored at -15℃~ -25℃ for one year.
- Yeasen kit has a high and stable library yield
Figure 1 Stable library yield
- Yeasen kit has a high mapping rate, a high proportion of exons, low Dup rate, a high number of genes detected, and low auto junction rate.
Figure 2 Excellent sequencing quality