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Published on: April 21, 2023
CODEC Library Preparation From Genomic DNA
James Phie1, Joshua N Johnstone1, Cameron Fraser1
1Systematic Medicine, Melbourne, Australia.
Current Protocols
|June 18, 2026
Summary
This study details a validated protocol for Concatenating Original Duplex for Error Correction (CODEC) library preparation. This method enhances DNA sequencing accuracy for detecting low-frequency somatic variants.
Area of Science:
- Molecular Biology
- Genomics
- Biotechnology
Background:
- Duplex sequencing offers enhanced accuracy by linking complementary DNA strands.
- Concatenating Original Duplex for Error Correction (CODEC) is a cost-effective duplex sequencing method.
- Widespread adoption of CODEC is hindered by a lack of detailed wet-lab protocols.
Purpose of the Study:
- To provide a detailed, experimentally validated protocol for CODEC library preparation.
- To ensure compatibility with standard Illumina sequencing platforms.
- To facilitate reproducible implementation of CODEC for research applications.
Main Methods:
- The protocol includes blunt-end fragmentation, dideoxy base-blocked A-tailing, and CODEC adapter ligation.
- Strand-displacing extension and library amplification are incorporated.
- Quality control checkpoints, critical parameters, and troubleshooting guidance are provided.
Main Results:
- An experimentally validated protocol for CODEC library preparation from genomic DNA is described.
- The protocol is compatible with standard Illumina sequencing platforms.
- An optional matched germline reference library preparation protocol is included.
Conclusions:
- This workflow enables reproducible CODEC library preparation in molecular biology labs.
- It supports accurate detection of low-frequency somatic variants.
- The protocol aims to broaden the adoption of cost-effective duplex sequencing.
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