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

•  High Exon Coverage Uniformity: Improved exon coverage uniformity enhances the accuracy of gene expression analysis.
  
• Premixed Reagents: Enzymes and buffers are pre-mixed to reduce reagent components and simplify preparation.
• Safe and Light-Resistant with Improved Strand Specificity: Optimized reverse transcriptase ensures high strand specificity without the need for light protection during handling.
• Preloaded Plate Format—Ready to Use: Designed for seamless integration with automation instruments, the preloaded plates enable easy “tear-and-use” operation for convenient automated workflows.

Components

[Note]: * The notation indicates that the component is not included in the kit. Primer mix is required if the complete adapters are used, otherwise it isn’t. This kit is compatible with Illumina and MGI platforms, but need additional primer mix (Cat # 13334 Primer Mix for MGI and Cat # 13335 Primer Mix for Illumina) for Illumina or MGI platform if the complete adapters are used.

** See the following diagram for the layout of plate reagent group.

Storage

This product should be stored at -25~-15℃ for one year..

Application

RNA library Preparation; Transcriptome profiling and gene expression analysis; mRNA sequencing for coding RNA studies; lncRNA (long non-coding RNA) research; Small RNA sequencing (if applicable); Differential gene expression analysis; RNA biomarker discovery; RNA variant and fusion detection.

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

Documents:

Safety Data Sheet

12340_MSDS_HB250620_EN.PDF

Manuals

12340_Manual_Ver.EN20251216.pdf

Related Blog:

A New Era of RNA Library Prep: The World’s First Pre-mixed Kit Without Toxic Actinomycin D