Product Description
Hieff NGS TM EvoMax RNA Library Prep Kit (Strand-specific) is a premixed, actinomycin D–free, strand-specific total RNA sequencing library preparation kit compatible with both Illumina and MGI platforms. Available in two formats—tube and sealing kit—this kit offers convenience for both automated liquid handling systems and manual operation. The kit includes reagents for RNA fragmentation, reverse transcription, strand-specific double-stranded cDNA synthesis, and library amplification. It can be combined with mRNA purification or rRNA removal kits for mRNA or lncRNA studies. By optimizing the reverse transcription step, this kit achieves high strand specificity without the use of actinomycin D, enhancing user safety. All reagents undergo stringent quality control and functional validation to ensure maximum stability and reproducibility in library preparation.
Features
Components
Storage
This product should be stored at -25~-15℃ for one year..
Applications
Figures
1.Simplified Product Format
Figure 1. Simplied components of EvoMax RNA Library Prep Kit. 2. High Exon Coverage Uniformity
Figure 2. Yeasen Kit show high exon coverage uniformity compared to NEB kit. The high GC exon were effectively detected. 3. High-Yield, Flexible RNA Library Preparation
Figure 3. Comparison of RNA Library Yields. (A, B) Strand-specific mRNA libraries from human, wheat, and mouse RNA (200 ng, 500 ng) using Hieff NGS™ EvoMax RNA Library Prep Kit (Strand-specific) (Cat# 12340ES) and mRNA Isolation Master Kit (Cat#12629) showed higher yield than Supplier N*. (A) Qubit quantification. (B) qPCR quantification. (C) Strand-specific LncRNA libraries from mouse RNA (200 ng, 500 ng) using the NGS Premix RNA Library Prep Kit with rRNA Depletion Kit (Human/Mouse/Rat, Cat#12726) showed higher yield than Supplier K* (qPCR). (D) Strand-specific LncRNA libraries from plant RNA (Arabidopsis, wheat, soybean leaves, 200 ng, 500 ng) using the NGS Premix RNA Library Prep Kit with rRNA Depletion Kit (Plant, Cat#12254) showed higher yield than Supplier K* (qPCR). 4. Higher Strand Specificity, More Transcripts
Figure 4. Comparison of mRNA Library Strand Specificity and Transcript Composition.
Figure 4. Comparison of mRNA Library Strand Specificity and Transcript Composition. (A) Human mRNA libraries prepared from 200 ng and 500 ng input demonstrated strand specificity of 99.4% and 99.3%(Cat#12340), compared with 99.0% and 99.2% for Supplier N*, respectively. (B) Transcript composition analysis showed that 12340ES libraries consistently detected more transcripts. With 200 ng input, 12340 detected 89.3% of coding + UTR transcripts versus 84.9% for Supplier N*. At 500 ng input, 12340ES detected 89.6% compared with 85.1% for Supplier N*. 5. Automation-Friendly
Figure 5. Comparison of Library Yields Between Automated and Manual Prep. (A).Four RNA standard samples (D5, D6, F7, M8) were used (3 replicates each) with the Hieff NGS™ EvoMax RNA Library Prep Kit (Strand-specific) (Cat# 12340ES) and mRNA Isolation Master Kit (Cat#12629). Libraries were prepared using the automated platform and manual workflow. Both automated and manual preps showed highly uniform library yields across all samples. (B). Accuracy of gene expression quantification. Fold-change analysis of RNA Library Prep Kit(Cat#12340)and mRNA Isolation Master Kit (Cat#12629)against the RNA standard dataset showed high consistency (Pearson r2 > 0.94). 6 Strong Stability
Figure 6. Stability Evaluation. (A) Accelerated stability was tested at 4°C and 25°C, with −20 °C storage as the control. (B) Real-time stability was monitored for up to 12 months. 7.RNA Library prep for different quality FFPE samples For severely degraded FFPE RNA (DV 200 <50%) and low input samples, we recommend a Double-purification protocol after adapter ligation to reduce library loss. Peak patterns of different quality FFPE samples FFPE RNA samples Sample 1 RIN=2.2; DV200=74% Sample 2 RIN=2.5; DV200=26% Sample 3 RIN=2.5; DV200=11% Sample 1. RNA Library Prep Results Input RNA: 500 ng, Fragmentation: 94℃, 7 min, Double purification : 0.6×; 0.8× Library amplification: 12 cycles, Library yield: 717.2ng Input RNA: 500 ng Fragmentation: 94℃, 7 min Purification & Selection: 0.6× & 0.7× / 0.15× Library amplification: 13 cycles Library yield: 437.8 ng Input RNA: 100 ng Fragmentation: 94℃, 7 min
Double purification: 0.6×; 0.8× Library amplification: 15 cycles Library yield: 206.8 ng Sample 2. RNA Library Prep Results Input RNA: 500 ng Fragmentation: 85℃, 8 min Double purification: 0.6×; 0.8× Library amplification: 12 cycles Library yield: 207 ng Input RNA: 500 ng Fragmentation: 85℃, 8 min Purification & Selection: 0.6× & 0.7× / 0.15× Library amplification: 13 cycles Library yield: 98.56 ng Sample 3. RNA Library Prep Results Input RNA: 500 ng Fragmentation: 65℃, 8 min Double purification: 0.6×; 0.8× Library amplification: 12 cycles Library yield: 354.2 ng Input amount: 500 ng Fragmentation: 65℃, 8 min Purification & Selection: 0.6× & 0.7× / 0.15× Library amplification: 13 cycles Library yield: 172.48 ng
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