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.
|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|
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.
Figure 1 Stable library yield
Figure 2 Excellent sequencing quality