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CORALL RNA-Seq V2 -

Your Key to Uncovering RNA Therapeutics Potential

Evaluate the activity of RNA therapeutics with CORALL RNA-Seq V2!

Time Saving

Time Saving

CORALL offers a fast and fully automatable whole transcriptome RNA-Seq workflow which is completed in only 4.5 hours.

Low Input Samples graphic

Excels with Low Input

CORALL supports the widest input range (1 ng - 1000 ng) with excellent performance for low input RNA down to 1 ng prior to mRNA-selection or ribo-depletion.

Adjustable Size

Adjustable Size

CORALL RNA-Seq V2 allows fragmentation-free library size adjustment to fit your application. Choose between medium size for expression analysis or longer lengths for annotation-based applications.

Upscale Experiments with Ease

Upscale Experiments with Ease

Automation-friendly and fast workflows allow processing of hundreds of samples in a single day and speed up turnover time to results!

Get the best sequencing results for demanding applications with CORALL RNA-Seq V2.


Add extra-confidence with built-in Unique Molecular Identifiers (UMIs) and process any sample type and quality with ease in only 6 steps: from sample to sequencing-ready library in less than one day.

Fast and flexible library prep in only 4.5 hours ⏱️

In contrast to conventional WTS RNA-Seq, CORALL library preparation requires only 6 steps and can be completed in 4.5 hours (Fig. 1).
CORALL does not require RNA fragmentation and is thus perfectly suited for processing of degraded or FFPE RNA.
 Figure 1 | Comparison of CORALL and conventional WTS RNA-Seq library preparation workflows

 Figure 1 | Comparison of CORALL and conventional WTS RNA-Seq library preparation workflows

CORALL Workflow

Removal of ribosomal RNA with RiboCop
Step 1: Removal of ribosomal RNA with RiboCop.

Samples are treated using a set of affinity probes for specific depletion of rRNA sequences and magnetic beads for depletion. RiboCop efficiently removes rRNA while maintaining unbiased transcript expression.
CORALL mRNA Workflow
Step 2:  CORALL Library Generation: RNA Input.
 
Ribo-depleted RNA can directly be used as template for CORALL reverse transcription. No prior RNA fragmentation is necessary, insert size is determined during reverse transcription.
CORALL total RNA-Seq V2 workflow
Step 2: CORALL Library Generation: Reverse Transcription.

CORALL library generation is initiated by random hybridization of Displacement Stop Primers (DSP) to the RNA template. The DSP also contains a partial Illumina adapter sequence. Reverse transcription extends each DSP to the next one, where transcription is effectively stopped. 
CORALL Total RNA-Seq V2 workflow

Step 2:  CORALL Library Generation: Linker Ligation.

Highly efficient ligation of Linker Oligos to the 3’ ends of first-strand cDNA fragments then introduces partial Illumina-adapters and Unique Molecular Identifiers (UMIs).

03_corall-mRNA-workflow
Step 3: CORALL Library Amplification: PCR.
 
During PCR, second strand synthesis is performed, and the double-stranded cDNA is amplified. Unique dual i7 and i5 indices and complete adapter sequences are introduced.
04_corall-mRNA-workflow
Step 4: Sequencing

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 complete workflow highly suitable for automation.

Transcriptome-wide evaluation of siRNA activity with CORALL

Small interfering RNAs (siRNAs) are recognized as RNA therapeutics with great potential against systemic diseases, such as cancer or infectious diseases.
 
RNA-Seq is the gold standard to evaluate the functional consequences of the treatment with RNA therapeutics such as siRNAs. Lexogen’s CORALL RNA-Seq is a fragmentation-free library preparation method with exceptional transcript start site coverage (Fig. 2). Therefore, CORALL is ideal to evaluate the effects of RNA therapeutics on coding and non-coding RNAs with transciption start site resolution.
 
Here, CORALL was used to confirm the anticancer activity of siRNAs against oncogenes in cervical cancer cells¹.
Figure 2 | Normalized ERCC coverage of transcription start sites

Figure 2 | Normalized ERCC coverage of transcription start sites. Normalized coverage of accumulated mapped reads for all detected ERCCs. The absolute nucleotide positions relative to the TSS (red dotted line) are shown.

Excellent start site representation

 
CORALL‘s comprehensive coverage delivers improved
transcription start site representation. Read coverage was analyzed using the ERCC spike-in controls, which feature precise, known transcription start sites (TSS).
 
Compared to competitor library preps using conventional fragmentation-based workflows, CORALL reads map more accurately to the exact ERCC transcription start sites (Fig. 2).
 
Hence, CORALL allows to interogate the effects of RNA therapeutics even on transcript isoforms that differ in their transcriptional start site usage.

Validation of anti-fibrotic siRNAs therapeutics

RNA interference (RNAi) is one of the antiviral defense mechanisms infected host cells employ. Thereby, virus-derived small interfering RNA (siRNA) are loaded onto host Argonaute proteins and trigger cleavage of the viral RNA via perfect sequence complementarity.
 
In this study, siRNAs were designed to serve as anti-COVID-19 therapeutic, targeting mRNAs of factors driving pulmonary fibrosis associated with a poor prognosis of COVID-19. CORALL was successfully used for differential transcript expression analysis to evaluate the impact of anti-fibrotic siRNA therapeutics².
Figure 3 | Exceptional performance on low input RNA.

Figure 3 | Exceptional performance on low input RNA. A) Excellent correlation for replicates of 1 ng Universal Human Reference RNA (UHRR) processed with CORALL mRNA-Seq V2. B) Read distribution for 1 ng input RNA for CORALL mRNA-Seq V2 and CORALL Total RNA-Seq V2 with RiboCop rRNA depletion.

Excellent performance on low input RNA

The complete CORALL RNA-Seq V2 workflow is highly reproducible (Fig. 3) and delivers high-quality RNA-Seq data even for low input samples down to 1 ng total RNA (prior to poly(A) selection or ribo-depletion).
 
CORALL is therefore highly suitable for analysis of precious samples with limited quantities or lowly concentrated RNA.

Wide input range and exceptional sensitivity

CORALL mRNA-Seq V2 displays exceptional gene discovery rates across the widest input range and is highly sensitive even for input as low as 1 ng total RNA prior to mRNA enrichment (Fig. 4). 
 
Add confidence to transcript quantification for low input amounts with CORALL's built-in UMIs.
Figure 4 | Gene discovery rates

Figure 4 | Gene discovery rates. CORALL mRNA-Seq libraries with long inserts were preprared from 1000 ng and 1 ng UHRR . The number of detected genes was plotted against the total number of uniquely mapping exonic reads.

Animated Gears

Upscale your experiments with ease!

CORALL RNA-Seq V2 library preparation workflows are fast and automation-friendly. Save time and process hundreds of samples in a single day.

CORALL RNA-Seq V2 Kits

CORALL RNA-Seq V2

Lexogen’s fast and flexible library prep kit with exceptional performance for challenging samples

  • Fragmentation-free RNA-Seq library prep with superior end-to-end coverage and true start site representation to cover all bases.

  • Built-in UMIs for highly accurate expression analysis 
    of coding and non-coding transcripts.

  •  CORALL RNA-Seq V2 supports the widest input range (1 ng - 1000 ng) with excellent sensitivity for low input RNA.

  • Easily scalable automation-friendly workflow.

Image of the CORALL v2 library prep kit from Lexogen, a set of reagents and protocols for preparing high-quality RNA sequencing libraries The kit enables efficient and reliable library preparation for a wide range of RNA sequencing applications..

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Ordering Information

  • Cat. №  | 171 - 176
    Product Name  CORALL RNA-Seq V2 Library Prep Kit with UDIs (Set A1, A2, A3, A4, B1, or A1- A4)
    Kit Sizes [preps] 24, 96, 384
     
  • Cat. №  | 177 - 182
    Product Name  CORALL mRNA-Seq V2 Library Prep Kit with UDIs (Set A1, A2, A3, A4, B1, or A1- A4)
    Kit Sizes [preps] 96, 384
     
  • Cat. №  | 183 -184
    Product Name  RiboCop HMR V2 and CORALL Total RNA-Seq V2 Library Prep Kit with UDIs (A1 or B1)   
    Kit Sizes [preps] 24, 96
     
  • Cat. №  | 185 - 186
    Product Name  RiboCop HMR+Globin and CORALL Total RNA-Seq V2 Library Prep Kit with UDIs (A1 or B1)     
    Kit Sizes [preps] 24, 96
     

Not sure which Kit to order?

Find the ideal kit for your application: 

References

1Gu et al., 2021, NAR (23): 1172-1190, doi: 10.1016/j.omtn.2021.01.018
2Ahn et al., 2022 Sci. Rep. (11): 19161, doi: 10.1038/s41598-021-98708-z

About Lexogen

Lexogen is a biotech company focusing on RNA and complete transcriptome studies using next generation sequencing technologies.

We have been at the forefront of RNA research since 2007, and our technologies and products are being used by thousands of scientists all over the world. “Lexo” in Greek means “word”, and Lexogen stands for the word of genes, which is the transcriptome. We strive to be the pioneers in this fascinating field of RNA science, and together with the fastest-growing technology of the past years, the next generation sequencing (NGS), we are sure that we can make a big contribution to the world.

Contact Us

Lexogen GmbH
Campus Vienna Biocenter 5
1030 Vienna, Austria

Lexogen, Inc.
51 Autumn Pond Park
Greenland, NH 03840, US

Email
info@lexogen.com

sales@lexogen.com

 Telephone:
US +1-603-431-4300
Rest of the world +43 (0) 1 345 1212 

 

Logo Iso certifications 9001 and 13485

Lexogen ISO certifications

We proudly hold both ISO 9001 and ISO 13485 certifications, which not only demonstrate our drive for innovation but also our unwavering devotion to delivering reliable, robust, and cutting-edge solutions to our valued customers.