Introduction to Illumina DNA library preparation

Next-generation sequencing (NGS) has become an essential tool across genetics, genomics, ecology, microbiology, agriculture, and biomedical research. Whether you're identifying genetic variants, sequencing microbial communities, assembling genomes, or analysing targeted regions, high-quality Illumina libraries are the foundation of reliable sequencing data.

Despite being one of the most widely used molecular techniques, DNA library preparation is often outsourced, leaving many students and researchers with little understanding of the process or the confidence to perform it themselves. In reality, library preparation is more accessible than many people expect, and developing this skill can significantly reduce project costs while providing greater flexibility for experimental design and troubleshooting.

This hands-on workshop will introduce participants to the principles of Illumina DNA library preparation for short-read sequencing before guiding them through the complete laboratory workflow. Participants will gain practical experience with the key steps involved in library construction, quality control, and best laboratory practices, while also learning how library preparation influences sequencing performance and downstream analyses.

Workshop aims

• Understand the principles behind Illumina short-read sequencing and DNA library preparation.
• Learn the complete workflow for preparing high-quality sequencing libraries.
• Gain hands-on experience with library preparation and quality control.
• Understand how library quality affects sequencing success and downstream analyses.
• Build the confidence to perform library preparation in your own laboratory.

Target audience: The workshop is designed for students and early career researchers with basic molecular biology laboratory experience who want to build practical genomics skills that can be immediately applied to their own research projects. If required, and based on registration responses, we will also offer an optional introductory laboratory session the day before the workshop to cover essential molecular biology techniques.

You are welcome to bring your own DNA samples to test whether this protocol is suitable for your organism. We will also provide samples during the workshop if needed. Please see the DNA quality-control requirements for samples.

Registration closes Monday 31 August. If you are planning on using your own DNA sample this should be checked and required information submitted by this date also.

Schedule

Day 1 — Sample preparation (9am, Tues 8th Sept)

• Pre lab introduction & theory session
• Sample prep workflow - sonicate, run gel, bead size selection, re-qubit

Day 2 — Library preparation (9am, Wed 9th Sept)

• Library prep workflow: end repair, adapter ligation, bead clean, PCR, bead clean

Day 3 — Results, discussion, and next steps (9am, Thur 10th Sept)

• Discuss results, troubleshooting, interpretation, next steps & bioinformatics

* please note that meal breaks and session end times will depend on lab work schedule and progress.