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Scalable Nucleic Acid Quality Assessments for Illumina NGS Library Prep
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2018-01-09 10:36:56
Scalable Nucleic Acid Quality Assessments for Illumina Next-Generation Sequencing Library Prep

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

Introduction

Assessing nucleic acid quality for library preparation is essential for the success of next-generation sequencing (NGS) applications. Verifying the integrity of nucleic acid samples before library preparation identifies samples likely to produce suboptimal libraries with poor sequencing performance due to degradation, fragmentation, or low purity. Similarly, the accurate assessment of both quality and quantity of prepared libraries can be used to optimize cluster generation during sequencing, resulting in cost- and timesavings while maximizing both sequencing data quality and output.

The Fragment Analyzer Automated Capillary Electrophoresis (CE) System from Advanced Analytical Technologies, Inc. (AATI) accelerates nucleic acid analysis workflows and provides a method that has been tested by Illumina for accurate quality assessment with multiple sample types. This application note provides an overview of the quality control (QC) workflow using the Fragment Analyzer during library preparation (Figure 1) and provides representative data plots for different sample and library types.

 

The Fragment Analyzer

The Fragment Analyzer is a proven solution for simultaneous qualification and quantification of DNA and RNA during library preparation for Illumina sequencing workflows. The Fragment Analyzer has many enhanced features, including:

* Diverse quantitative sample kit options for genomic DNA (gDNA), NGS libraries, small RNA, total RNA, and messenger RNA (mRNA)

* Seamless, automated switching between applications with two gel input lines

* Scalable array options for parallel CE analysis of 12, 48, or 96 samples

* Multitray format holds three standard 96-well plates for automated analysis of up to 288 samples

* Minimal hands-on time required for instrument setup and sample handling

* High analytical sensitivity detects concentrations as low as 5 pg/µl for fragments and 50 pg/µl for smears

* High throughput support with the Fragment Analyzer INFINITY ® Automated CE System, designed to integrate with robotic cells for automated, continuous operation, capable of running over 2400 samples daily without user intervention

 

Figure 1: Illumina NGS Library Prep QC Workflow—The Fragment Analyzer is used to assess the quality and quantity of nucleic acids after isolation and after library preparation to enable generation of high-quality sequencing data.

 

 

 

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