CORALL - mRNA-Seq Library Prep Kit

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).

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.

A Data Analysis Pipeline is now available on the BlueBee® Genomics Platform. The provided pipeline enables kit users to perform read quality control, mapping, Unique Molecular Identifier (UMI) deduplication, and transcript quantification. For using your activation code register an account with BlueBee (https://lexogen.bluebee.com/portal) and upload your data (fastq.gz files).

NOTE! BlueBee Data Analysis is available for a range of species, please refer to FAQ 7. Reference genomes for new species can be added upon request. Please note this will incur a fee. See CORALL Data Analysis, for further details.

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.

Fast and Easy All-in-One Workflow

CORALL mRNA-Seq Kits provide a complete solution for mRNA sequencing. RNA is poly(A) enriched in 1.1 hours and can be directly transferred into CORALL library preps. The streamlined RNA fragmentation-free protocol generates ready-to-sequence libraries in 4.5 hours. The complete CORALL mRNA-Seq workflow takes less than 6 hours.