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CORALL  Total RNA-Seq V1

The CORALL Total RNA-Seq Library Prep Kit enables fast and cost-efficient generation of UMI labelled, stranded libraries for whole transcriptome 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 generates transcriptome-wide smooth, uniform read coverage, comparable to that of competitor kits (Fig. 1).

CORALL_Figure_01

Figure 1 | 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 delivers excellent gene discovery rates matching conventional competitor kits (Fig. 2A). 95% of genes are commonly detected between CORALL and each of the two competitors (at >10 counts per million (CPM), Fig. 2B). This high level of library complexity ensures faithful representation of the transcriptome, enabling sensitive expression profiling.

CORALL_Figure_02_A_B

Figure 2 | Gene detection. A) Gene discovery rates. The number of detected genes is plotted against the total number of reads mapping uniquely to exons (calculated with featureCounts). B) Overlap of detected genes. The Venn diagram illustrates overlaps between CORALL and competitor kits for genes detected with normalized expression levels >10 CPM (for uniquely mapping reads).

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 libraries can be prepared from poly(A)-enriched or rRNA-depleted RNA (100 – 200 ng of total RNA yields approximately 1 – 2 ng of rRNA-depleted RNA). CORALL is available as a stand-alone kit with single indexing or UDIs (Cat. No. 095, or 117 – 119 and 132 – 134) or in a bundle with RiboCop rRNA Depletion Kits (Cat. No. 146, 147). For mRNA analysis we recommend to use the Poly(A) RNA Selection Kit (Cat. No. 039) upstream of CORALL. Total RNA without prior depletion or enrichment (100 pg to 100 ng) can also be used as input.

UMIs are seamlessly introduced into CORALL libraries as an inherent part of the protocol. As a result 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.

Submit your CORALL data analysis request.

CORALL libraries can be multiplexed using up to 96 i7 indices (included in the single indexing kit, Cat. No. 095, 146). Additional i5 indices can also be introduced using the Lexogen i5 6 nt Dual Indexing Add-on Kits (Cat. No. 047). Used together, Lexogen’s 96 i7 and 96 i5 6 nt indices enable up to 9,216 different index combinations for sequencing.

NEW! CORALL Total RNA-Seq library prep is now also available with up to 384 pre-mixed Unique Dual Indices (Cat. No. 117-119, 132-134). UDIs are also included in CORALL mRNA-Seq Kits (Cat. No. 158-163). 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.

The streamlined RNA fragmentation-free protocol generates ready-to-sequence libraries in 4.5 hours.

“We have successfully used the CORALL kit in our latest project to generate high quality sequencing libraries from low input material. With its fast and clear protocol we were able to generate the libraries in less than one day. We will surely use the CORALL kit again and recommend it for whole transcriptome analysis.”

Jason Sims, PostDoc, Schlögelhofer Group, Department of Chromosome Biology, Max Perutz Labs, Vienna

“We used the new CORALL kit for performing transcriptome-analysis of CRISPR-modified cells in order to understand the consequences of deregulated epigenetic modifiers. In our hands the kit performance was highly satisfying in terms of data-quality and reproducibility across biological replicates. It furthermore convinced us with the ease of use, clarity of instructions, details in the manual, and handling of reagents.”

Max Koeppel, Head of Functional Tumor Genomics Group at Leibniz-Institute DSMZ, Germany

CORALL kit has been key for our latest project. Despite having extremely heterogeneous samples, we were able to produce successful libraries from, literally, all our samples on the first attempt. Its perfect integration with Ribocop rRNA depletion kit and compatibility with SIRVs has made the project more straightforward than what we anticipated. Finally, as we required UMIs for demultiplexing, CORALL ticked all the boxes. We will use it again for our future projects.”

Eneko Villanueva Verdejo, Cambridge Centre for Proteomics, Department of Biochemistry, University of Cambridge, Cambridge, UK

Workflow

Reverse Transcription
Step 1:
CORALL Total RNA-Seq Library Prep uses total,
poly(A) enriched or rRNA depleted RNA as input.

corall_workflow_01_V2timer_corall_v2

Reverse Transcription
Step 1:
CORALL library generation is initiated by random
hybridization of Displacement Stop Primers (DSP)
with partial Illumina-compatible P7 sequences,
to the RNA template. No prior RNA fragmentation is
necessary, as the insert size is determined by the
distance between hybridized DSPs. Reverse transcription
extends each DSP to the next, where transcription is
effectively stopped. This stop prevents spurious second
strand synthesis, maintaining excellent strand specificity.

corall_workflow_02_V2timer_corall_v2

Linker Ligation
Step 2:
Highly efficient ligation of Linker Oligos to the 3’ ends
of first-strand cDNA fragments then introduces
partial Illumina-compatible P5 sequences
and Unique Molecular Identifiers (UMIs).

timer_corall_v2

Library Amplification
Step 3:
During PCR, second strand synthesis is performed,
and the double-stranded cDNA is amplified. In doing so,
i7 and (optional) i5 indices as well as complete adapter
sequences required for cluster generation on Illumina
instruments are added.

timer_corall_v2

Sequencing
Step 4:
The UMI is positioned to be read out at the beginning
of Read 1 during sequencing and can therefore be
assessed even in Single-Read mode. All purification
steps are based on magnetic beads, rendering
the protocol highly suitable for automation.

corall_workflow_05_V2timer_corall_v2

FAQ

Frequently Asked Questions

Access our frequently asked question (FAQ) resources via the buttons below.

Please also check our General Guidelines and FAQ resources!

How do you like the new online FAQ resource? Please share your feedback with us!

Downloads

CORALL Total RNA-Seq Library Prep Kit for Illumina

pdf User Guide for CORALL Total RNA-Seq – update 31.03.2020
pdf User Guide for CORALL RNA-Seq with UDIs – update 28.03.2023
pdf User Guide for Lexogen 12 nt Unique Dual Indexing Add-on Kit – update 25.01.2023
pdf Product Flyer – update 20.09.2019
pdf PCR Add-on Kit for Illumina User Guide – update 03.01.2023
pdf Lexogen i5 6 nt Dual Indexing Add-on Kits (5001-5096) User Guide – update 03.01.2023

pdf Lexogen UDI 12 nt Unique Dual Index Sequences – update 26.03.2021
pdf Lexogen i7 and i5 Index Sequences – update 05.05.2020

 Library Quantification File
Demultiplexing and Error Correction Tool – iDemux

Safety Data Sheet

Request Safety Data Sheet

If you need more information about our products, please contact us through support@lexogen.com or directly under +43 1 345 1212-41.

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