Multi-Platform, Multi-Method RNA Library Preparation
RNA is a single strand formed by transcribing one strand of DNA as a template, following the principle of complementary base pairing. Its primary function is to realize the expression of genetic information in proteins, serving as a bridge in the transmission of genetic information. It mainly exists in biological cells as well as some viruses and viroid.
As is widely known, more than 95% of total RNA consists of ribosomal RNA (rRNA), yet rRNA is highly conserved and exhibits extremely stable expression across different tissues and organs. In contrast, messenger RNA (mRNA), which accounts for only 2% to 3% of total RNA, contains the most abundant informational content among RNAs. Therefore, mRNA has become a major focus in scientific research.
Pie Chart of Cellular RNA Distribution
The transcription of mRNA products in specific tissues or cells of a particular species under a given physiological functional state, known as the transcriptome, represents a critical step and process in the transmission and expression of genetic information in the genome.
Through high-throughput mRNA sequencing, vast amounts of transcript sequence information can be obtained, enabling the quantitative assessment of gene transcription expression levels and the acquisition of genomic transcription regions and their loci. This plays a significant role in genome sequence assembly and annotation, the study of differential gene expression across samples, and functional research. It allows for the investigation of gene function and structure at a holistic level, revealing the molecular mechanisms underlying specific biological processes and disease development. This approach has been widely applied in basic research, clinical diagnosis, and drug development.
The quality of RNA sample library preparation is crucial for subsequent sequencing, as it can directly impact the analysis of results. Therefore, a reliable library preparation kit is essential for RNA sequencing. TransGen Biotech has currently launched two RNA library preparation kits, compatible with Illumina and MGI platforms respectively, both characterized by high data quality and high library conversion rates.
Fast RNA library prep
➣ TransNGS® Fast RNA-Seq Library Prep Kit for Illumina®
This kit is specifically designed for the Illumina high-throughput sequencing platform to construct either strand-specific or non-strand-specific transcriptome libraries. It enables one-step completion of second-strand synthesis, end repair, and A-tailing reactions without requiring intermediate purification steps, significantly simplifying the operational workflow and reducing library preparation time. Within just 3 hours, it can construct high-quality sequencing libraries from 0.1 ng to 100 ng of processed RNA, yielding libraries with high output and intact information.
Comparison Data
Purify mRNA from total RNA of human, mouse, wheat, and tobacco using an mRNA purification kit, with input amounts of 200 ng total RNA for human and mouse, and 1000 ng total RNA for wheat and tobacco. Subsequently, construct RNA libraries using library preparation kits from TransGen and Company V, respectively. After library size selection, evaluate library yield and profile, and perform sequencing analysis.
Library Yield
Using high-quality RNA samples (RIN > 8) from nine different species, we perform mRNA capture and subsequent library preparation. The results demonstrate that TransGen's strand-specific and non-strand-specific library preparation kits consistently generated higher library yields compared to those of Company Y.
Using severely degraded RNA (RIN = 3) from human (293T cells), mouse, and tobacco, we perform mRNA capture followed by library construction with high (1 μg) and low (200 ng) input amounts. The results demonstrate that the TransGen's kit (both strand-specific and non-strand-specific) consistently outperforms Company Y in library yield across all species and input amounts.
Sequencing Data Quality
Comparative analysis of sequencing data across multiple species reveal that RNA libraries using TransGen kit significantly outperform Company Y’s in percentage of clean data, number of reliably detected genes.
➣ TransNGS® Fast RNA-Seq Library Prep Kit for MGI®
This kit is specifically designed for the MGI high-throughput sequencing platform to construct either strand-specific or non-strand-specific transcriptome libraries. It enables one-step completion of second-strand synthesis, end repair, and A-tailing reactions without requiring intermediate purification steps, significantly simplifying the operational workflow and reducing library preparation time. Within just 3 hours, it can construct high-quality sequencing libraries from 0.1 ng to 100 ng of processed RNA, yielding libraries with high output and intact information.
Schematic Diagram of Library Preparation
Comparison Data
The RNA library construction performance is compared between TransGen and Company Y’s products using RNA samples with different input amounts, varying integrity levels and from various species. The TransGen’s products demonstrate significantly higher yields compared to competitors, achieve over 98% strand specificity across all types of strand-specific libraries, and consistently produce high-quality sequencing data.
Library Yield
Library Profile
Sequencing Data Quality
Accessories
➣ TransNGS® rRNA Depletion Kit (Human/Mouse/Rat)
Features
• Remove up to 99% of rRNA in humans/mouse/rats effectively .
• Provide Control qPCR Primer Sets which are able to detect changes in rRNA and non-ribosomal RNA content before and after removal.
Stability of the Product
Different batches of the product are used to remove rRNA from 1 μg of total RNA (HepG2 cells). Both untreated and rRNA-depleted RNA samples are then analyzed by qRT-PCR using rRNA-specific primers and non-rRNA primers. The stability between product batches is assessed based on the ΔCq value (Cq value of the depleted sample minus the Cq value of the untreated sample). No significant differences in ΔCq values are observed between different batches, indicating good batch-to-batch consistency.
Comparison Data
For 1 μg of total RNA samples from human (H), mouse (M), and rat (R), rRNA is removed using products from TransGen and Company K. The rRNA-depleted RNA is then used for library preparation (TransGen, KP701). The results demonstrate that TransGen's product performs comparably to—and in some aspects even outperforms—the competitor's product in terms of sequencing data quality.
Sequencing Data Quality
Features
• High mRNA capture efficiency
• Excellent sample compatibility
• High-quality sequencing data
• Compatible with a wide range of species
The TransGen mRNA Purification Kit (EC511) and comparable products from Company V and Company Y are used to capture mRNA from varying input amounts and different species for library preparation. The results demonstrate that TransGen's product delivers higher mRNA yield and purity. When paired with library prep kits, it generates higher library yields and superior sequencing data quality compared to competitor’s products.
Product Information
