Paired DNA and RNA profiling is increasingly utilized in genomics research to investigate correlations between personal genotype and phenotype and unveil molecular mechanisms of disease.
However, parallel or sequential library preparation for paired sequencing can be time-consuming, require large sample volumes and produce variable results.
This infographic showcases a new combined, single-tube prep workflow that simultaneously generates DNA and RNA libraries from a single input formalin-fixed paraffin-embedded (FFPE) sample.
Download this infographic to discover how to:
- Save time and resources to accelerate paired sequencing preparation
- Conserve sample and reduce bias while detecting both genomic variants and expressed transcripts
- Detect DNA and RNA biomarkers from the same total nucleic acid input
KAPA Total Prep FFPE workflow KAPA Total Prep FFPE workflow simultaneously generates DNA and RNA libraries from a single input FFPE sample. Advantages over separate DNA and RNA library prep: Saves time, resources, and labware Conserves often-limited FFPE sample Enables more reliable tumor profiling by reducing bias caused by FFPE sample variability Save time and resources compared to parallel or sequential library preparation Total Nucleic Acid NGS Library Prep Tech Spotlight Get more out of your paired sequencing with a combined, single-tube library prep for DNA and RNA Simultaneously capture regions of interest from the genome and corresponding transcripts with optional target enrichment ATCGGCA CGCATCG GACTACG TTTGGCC ATCGGCA CGCATCG GACTACG TTTGGCC ATCGGCA CGCATCG GACTACG TTTGGCC ATCGGCA CGCATCG GACTACG TTTGGCC Typical workflow for paired sequencing More total time More hands-on time More sample needed More variability and bias Less total time Less hands-on time Less sample needed Less variability and bias ATCGGCA CGCATCG GACTACG TTTGGCC ATCGGCA CGCATCG GACTACG TTTGGCC ATCGGCA CGCATCG GACTACG TTTGGCC ATCGGCA CGCATCG GACTACG TTTGGCC Sample 1 DNA Library preparation NGS sequencing Sample 2 RNA Library preparation NGS sequencing Data analysis Sample Total Nucleic Acid (DNA + RNA) Single-tube library preparation Data analysis NGS sequencing New KAPA Total Prep Workflow RNA molecules are tagged to enable sorting of RNA and DNA reads during analysis If target enrichment is included, there is no need for rRNA depletion Target enrichment enables more focused sequencing while reducing sequencing costs Sample Total Nucleic Acid (DNA + RNA) Single-tube library preparation Data analysis NGS sequencing Optional target enrichment New KAPA Total Prep WorkflowMC-US-13724 A777 09/23 Project: Single-Tube Total Nucleic Acid Library Preparation for FFPE, Wilmington, MA 2023 For Research Use Only. Not for use in diagnostic procedures. KAPA and HYPERCAP are trademarks of Roche. All other product names and trademarks are the property of their respective owners. © 2023 Roche Sequencing and Life Science. All rights reserved. Learn more about the KAPA Total Prep FFPE workflow and how it can help you at go.roche.com/TotalPrep or by scanning the QR code. SNV and InDel allele frequency in the DNA reads is concordant between manual and KAPA Total Prep library preparation workflows. Both workflows included target enrichment with the KAPA HyperCap Oncology Panel. The KAPA Total Prep workflow enables the detection of RNA fusions and exon-skipping events. Detection of 16 RNA known fusions and 2 exon-skipping events was analyzed with the KAPA Total Prep FFPE workflow workflow. SNV allele frequency is consistent between DNA and RNA reads. SNVs were detected in an FFPE sample with known expected allele frequencies using the KAPA Total Prep FFPE workflow, with the KAPA HyperCap Oncology Panel. Results for both DNA and RNA reads were compared to the expected allele frequencies. For Research Use Only. Not for use in diagnostic procedures. | © 2023 Roche 2 SNV allele frequency is consistent between DNA and RNA reads Expected DNA reads RNA reads Allele frequency (%) For Research Use Only. Not for use in diagnostic procedures. | © 2023 Roche 1 SNV and InDel allele frequency in the DNA reads is concordant between library preparation workflows KAPA Total Prep workflow Standard DNA library preparation R2: 0.863 Allele frequency (%) For Research Use Only. Not for use in diagnostic procedures. | © 2023 Roche The Single-tube TNA Workflow enables the detection of RNA fusions Gene fusions Exon skipping 93.8% detected 100% detected Conserve sample while detecting both genomic variants and expressed transcripts Detect DNA & RNA biomarkers from the same total nucleic acid input SNVs InDels CNVs Gene fusions Exon skipping events Splicing isoforms Gene expression levels For Research Use Only. Not for use in diagnostic procedures. | © 2
