DNA PCR-free library construction using fully automated sample preparation

Next generation sequencing (NGS) is a powerful technology that requires reliable and cost-effective protocols for the construction of DNA libraries. Increased accessibility to NGS platforms, the growth of sequencing applications and reduced sequencing costs has fueled an increasing demand for higher-throughput sample preparation.

Manual sample preparation for DNA library construction is a multi-step process that can be time-consuming, tedious and prone to human error. Many of the steps involved can be automated to standardize sample preparation, increase efficiency and reduce costs. Laboratories that adopt automated sample preparation often report reductions in hands-on time, a higher throughput and a greater yield of sequence-ready libraries, compared with manual sample preparation.

Beckman Coulter scientists have now introduced a protocol for automating Illumina’s TruSeq® DNA PCR-Free library construction kit on the Beckman Coulter Biomek® FXP Liquid Handler. This method generates libraries that do not undergo PCR amplification, which reduces nucleic acid composition bias and eliminates the risk of artifacts being introduced. The Biomek automated TruSeq DNA PCR-Free method offers full automation of DNA sample preparation, with each method run processing up to 96 samples simultaneously in approximately five hours.

Using this process, users can choose to perform incubation off-line using an external bench top thermal cycler machine or on-deck using an integrated TRobot thermal cycler. The Biomek FXP is configured with Span 8 probes for the transfer of reagents and enzymes and a 96-multichannel pipetting head for DNA clean up, wash and elution transfers. A static Peltier unit keeps enzyme master mixes cool throughout the method run.

As described in a recent application note, Beckman Coulter assessed the construction, quality, size distribution and concentration of the libraries created using the Biomek automated TruSeq DNA PCR-Free method. The results showed a yield of amplification-free, sequence-ready libraries that was more than twice the minimum amount needed for the Illumina TruSeq DNA PCR-Free protocol.

The Biomek automated TruSeq DNA PCR-Free method for library sample preparation provides a highly efficient, high throughput and cost-effective automation solution for DNA library construction. Compared to manual sample preparation, this method significantly decreases human error rate and increases yield, while dramatically reducing the amount of hands-on time researchers have to spend on preparation steps.

To find out more about the Biomek automated TruSeq DNA PCR-Free method for DNA library construction, please click here.

Beckman Coulter Life Sciences - Auto-Cellular and ProteomicsAbout Beckman Coulter

Beckman Coulter develops, manufactures and markets products that simplify, automate and innovate complex biomedical tests. More than a quarter of a million Beckman Coulter instruments operate in laboratories around the world, supplying critical information for improving patient health and reducing the cost of care.


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Last updated: Mar 23, 2020 at 12:38 PM

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