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Illumina sequencing library preparation for highly multiplexed target capture and sequencing
Matthias Meyer1, Martin Kircher
1Max Planck Institute for Evolutionary Anthropology, D-04103 Leipzig, Germany. mmeyer@eva.mpg.de
Cold Spring Harbor Protocols
|June 3, 2010
Summary
This study presents a fast, reliable, and cost-effective protocol for preparing barcoded DNA sequencing libraries. This method enables parallel sequencing of multiple samples, essential for high-throughput genomics research.
Area of Science:
- Genomics
- Molecular Biology
- Bioinformatics
Background:
- High-throughput sequencing generates vast DNA data, enabling parallel sample analysis.
- Target capture is now preferred over polymerase chain reaction (PCR) for target enrichment.
- Parallel sequencing necessitates sample-specific barcodes for accurate source tracing.
Purpose of the Study:
- To describe a rapid and dependable protocol for preparing indexed sequencing libraries.
- To facilitate parallel target capture and sequencing of multiple samples.
- To offer a cost-effective alternative to commercial library preparation kits.
Main Methods:
- Development of a protocol for barcoded (indexed) sequencing library preparation.
- Utilizes a 96-well plate format suitable for multi-channel pipettes.
- Adaptable for various double-stranded DNA inputs, including subnanogram quantities.
Main Results:
- The protocol is fast, completed within two to three days.
- It is reliable and cost-effective, avoiding expensive commercial kits.
- Successfully prepares indexed libraries for Illumina's Genome Analyzer platform.
Conclusions:
- This protocol provides an efficient method for preparing barcoded sequencing libraries.
- It supports parallel sequencing of multiple samples, crucial for large-scale genomics.
- The method is accessible and cost-effective for researchers.
