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 %).
Superior End-to-End Coverage
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.
Unique Molecular Identifiers (UMIs) seamlessly included
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. For further questions, please contact firstname.lastname@example.org.
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.
“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.”
Poly(A) RNA is enriched by hybridization to oligo(dT)
magnetic beads. Any RNA without poly(A) stretches
is washed away and poly(A) RNA is finally eluted.
for CORALL reverse transcription. No prior RNA fragmentation
is necessary, insert size is determined during reverse transcription.
of Displacement Stop Primers (DSP) with partial Illumina-compatible
P7 sequences, to the RNA template. Reverse transcription extends
each DSP to the next one, where transcription is effectively stopped.
This stop prevents spurious second strand synthesis, maintaining
excellent strand specificity.
cDNA fragments then introduces partial Illumina-compatible P5
sequences and Unique Molecular Identifiers (UMIs).
and the double-stranded cDNA is amplified. In doing so,
unique dual i7 and i5 indices as well as complete adapter
sequences required for cluster generation on Illumina
instruments are added.
during sequencing and can therefore be assessed even in Single-Read
mode. All purification steps are based on magnetic beads, rendering
the complete workflow highly suitable for automation.
Frequently Asked Questions
Please find a list of the most frequently asked questions below. If you cannot find the answer to your question here or want to know more about our products, please contact email@example.com.
CORALL mRNA-Seq Library Prep Kit
User Guide – CORALL RNA-Seq Integrated Data Analysis Pipelines on BlueBee® Genomics Platform – update 20.01.2021
PCR Add-on Kit for Illumina Instruction Manual – update 04.11.2020
Lexogen UDI 12 nt Unique Dual Index Sequences – update 26.03.2021
Material Safety Datasheets
If you need more information about our products, please contact us through firstname.lastname@example.org or directly under +43 1 345 1212-41.
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