CORALL mRNA-Seq V1
The CORALL mRNA-Seq Library Prep Kit enables fast and cost-efficient generation of stranded, UMI labelled, and unique dual indexed libraries for whole transcriptome poly(A) RNA analyses using Illumina® NGS platforms.
Dear Customer, CORALL V1 kits will be phased out soon. The last order date is October 31, 2023.
For more information, please get in touch with us at info@lexogen.com or sales@lexogen.com. Thank you!
Performance
CORALL delivers excellent gene discovery rates across a wide range of RNA input amounts (Fig. 1).
Figure 1 | Gene discovery rates. 100 ng, 10 ng, and 1 ng Universal Human Reference RNA (UHRR) were used as input for poly(A) selection and library preparation using CORALL mRNA-Seq. Libraries were sequenced on Illumina® NextSeq500 (1×150 bp, 11.2 M reads / sample). The number of detected genes is plotted against the number of reads mapping uniquely to exons (calculated with featureCounts).
CORALL generates transcriptome-wide smooth, uniform read coverage, comparable to that of competitor kits (Fig. 2).
Figure 2 | Accumulated transcript body coverage (whole transcriptome). Coverage across all transcripts was generated using the geneBody_coverage.py tool provided by RSeQC (transcripts length normalized to 100 %).
CORALL’s comprehensive coverage delivers improved transcript start and end site representation. Read coverage was analyzed using the ERCC spike-in controls, which feature precise, known transcription start and end sites (TSS and TES, respectively). CORALL reads map more accurately to the exact ERCC TSS (Fig. 3A) than competitor libraries, which fail to cover the true start sites. Additionally, CORALL provides elevated coverage at TES (Fig. 3B).
Figure 3 | Normalized ERCC coverage of A) TSS and B) TES. Normalized coverage of accumulated mapped reads for all detected ERCCs. The absolute nucleotide positions relative to the TSS (red dotted line, A) and TES (blue dotted line, B) are shown.
CORALL mRNA-Seq Kits are the optimal choice for whole transcriptome mRNA sequencing. The kits include Poly(A) Selection for mRNA-enrichment prior to CORALL library generation and Lexogen’s UDI 12 nt Unique Dual Index Sets for amplification of unique dual indexed libraries. CORALL mRNA-Seq is compatible with total RNA inputs from 1 ng – 1 µg. High quality RNA (RIN, RQN >8) is recommended. CORALL libraries can also be prepared from rRNA-depleted RNA, e.g., using Lexogen’s RiboCop rRNA Depletion Kits. For samples with RNA integrity or quality scores (RIN, RQN) below 8, the CORALL Total RNA-Seq workflow with rRNA-depletion is recommended instead of CORALL mRNA-Seq.
UMIs are seamlessly introduced into CORALL libraries allowing to remove PCR duplicates for accurate quantification. Read 1 contains the UMI information making it directly accessible in the cost-efficient single-read mode.
Lexogen offers an automated solution for CORALL data analysis based on our state-of-the-art proprietary pipeline, which allows researchers to analyze CORALL samples in a convenient and fast way.
Codes for CORALL data analysis on Lexogen’s automated data analysis solution are available for purchase. Please contact us at sales@lexogen.com.
CORALL mRNA-Seq Kits are available with up to 384 pre-mixed Unique Dual Indices (UDIs) for Forward Strand (A) and Reverse Complement (B) workflow (Set A1-A4: Cat. No. 158-161, 163, or Set B1: Cat. No. 162). Lexogen’s new 12 nt UDIs feature superior error correction for maximal sequencing data output and are also available as stand-alone Add-on Kits (Cat. No. 107 – 111) for use with other library preps. For further questions, please contact support@lexogen.com.
“Transcriptional profiling is one of our key approaches to study and understand the molecular mechanism of action of cancer drugs. We have made excellent experiences with the CORALL kit both in terms of experimental feasibility but also data quality. Through integration with synthetic spike-ins SIRVs, It was particularly helpful when testing drugs that affect overall transcriptional output and thus cause a global decrease in mRNA levels.”
Workflow
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FAQ
Frequently Asked Questions
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Please also check our General Guidelines and FAQ resources!
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Downloads
CORALL mRNA-Seq Library Prep Kit
User Guide for Poly(A) Selection Kit – update 13.12.2022
User Guide for CORALL RNA-Seq with UDIs – update 28.03.2023
PCR Add-on Kit for Illumina User Guide – update 03.01.2023
Lexogen UDI 12 nt Unique Dual Index Sequences – update 26.03.2021
Library Quantification File
Demultiplexing and Error Correction Tool – iDemux
Safety Data Sheet
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