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NGS Library Preparation

What is NGS Library Preparation

What is Library Preparation?

Next-generation sequencing (NGS) library preparation involves generating a collection of DNA fragments for sequencing. NGS libraries are typically prepared by fragmenting a genomic DNA or cDNA sample and ligating specialized adapters to both fragment ends. An alternative process called "tagmentation" combines the fragmentation and ligation reactions into a single step. Adapter-ligated fragments are then PCR-amplified and gel-purified.

Illumina Library Preparation Highlights

High-quality library preparation is critical for a successful NGS experiment. Illumina offers optimized sequencing library preparation solutions for a wide variety of NGS methods, including whole-genome sequencing, targeted DNA sequencing, whole-transcriptome sequencing, targeted RNA sequencing, and more.

Illumina library prep protocols can accommodate a range of throughput needs, from lower-throughput protocols for small laboratories to fully automated library preparation workstations for large laboratories or genome centers. Our solutions support a broad range of sample types, from cell lines to fresh tissue, formalin-fixed paraffin-embedded (FFPE) samples, blood, and other challenging sample types.

Kits for Preparing NGS Libraries

Kits for NGS Library Preparation

Illumina offers an extensive assortment of easy-to-use next-generation sequencing library preparation kits for DNA, RNA, and epigenetic sequencing studies. To find the right kit for your needs, use the selection tool below, or view a filterable list of library prep kits.

Library Prep Kit Selector

High-Throughput Automation Solutions

For labs preparing large quantities of NGS libraries, liquid-handling robots and additional automation solutions provide high-throughput capacity and important quality control touch points. Visit the page below to learn more about Illumina-qualified sequencing library prep automation methods available through our partners.

See Automation Options
Automated Library Prep Buyer's Guide

This guide will walk you through the range of automation tools that exist for NGS library preparation, and help you find the right solution for your needs.

Read Buyer's Guide
NGS Workflow Steps

Learn the basics of each step in the Illumina sequencing workflow and find out how to prepare before starting the library preparation process.

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Featured Microbial Whole-Genome Sequencing Workflow

Library Prep Technical Tips

DNA/RNA Isolation Considerations

Find guidance to help you avoid contamination while purifying DNA/RNA before library preparation.

Read Technical Bulletin

dsDNA Library Concentration

Learn how to convert library concentration from ng/┬Ál to nM for some Illumina library preparation methods.

Read Technical Bulletin

Quantification and Quality Control

Find out how to quantify and validate final libraries for a successful sequencing run.

Read Technical Bulletin

Library Normalization Best Practices

Learn when library normalization is required, and how to perform the necessary steps.

Read Technical Bulletin

What is the PhiX Control v3 Library?

Learn how to use the PhiX library as a control in your Illumina sequencing runs.

Read Technical Bulletin

Nucleic Acid Quality Assessment

Simultaneous qualification and quantification of nucleic acids with the Fragment Analyzer.

Read Technical Bulletin

Sequencing Library Preparation Methods

Access a summary of NGS methods compiled from peer-reviewed publications. Explore the wide range of questions that can be addressed with Illumina technology. Find method descriptions as well as pros and cons.

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Methods GuideAccess Guide
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Additional Resources

Beginner's Guide to Next-Generation Sequencing
Beginner's Guide to Next-Generation Sequencing

In this introduction, we'll guide you through the steps in the NGS workflow and planning your first experiment.

Push-Button Informatics
Push-Button Informatics

Illumina bioinformatics tools make it easier to manage, analyze, and interpret large amounts of complex genomic data.

Multiplex Sequencing and Index Adapters
Multiplex Sequencing and Index Adapters

Find out more about sample multiplexing and compare the different types of index adapters.