Integrated NGS library prep and quantification in a single workflow

NuQuant®: A novel method to accurately measure molar concentrations of NGS libraries in seconds

The NuQuant library quantification method accurately measures molar library concentration without the need for separate fragment size analysis. NuQuant library quantification is a proprietary method by which a specific number of fluorescent labels are incorporated into the library molecules during library preparation. Consequently each library molecule has an equivalent number of labels incorporated, regardless of the size of the library fragment. The library molar concentration can be directly measured using fluorometers like Qubit® or standard plate readers.

NuQuant Library quantification is integrated in Celero DNA-Seq and Universal Plus mRNA-Seq with NuQuant library preparation kits.

Why Use NuQuant?

Accurate quantification of Next-Generation Sequencing (NGS) libraries for Illumina sequencers is critical to ensure efficient use of sequencing capacity, avoid over-clustering or under-clustering of the flow cell and generate sufficient reads for each sample in a multiplexed library. Current methods for Illumina® library quantification such as quantitative PCR (qPCR), fluorometric mass measurement with Qubit or microfluidic electrophoresis with Bioanalyzer® are time consuming and require multiple steps. qPCR needs a long preparation time with several dilution steps that can introduce variability. Libary quantification with qPCR also requires expensive reagents and standards, and secondary library analysis methods such as Bioanalyzer to determine average library size. NuQuant library quantification provides library molar concentration with a simple fluorescent measurement that does not require serial dilutions, additional reagents or secondary library analysis methods.

Some of the unique benefits of quantifying NGS library concentraion with NuQuant include:

  • Quantify molar concentration of libraries without the Bioanalyzer step unlike other methods like qPCR and Qubit 
  • Save valuable time and resources since NuQuant library quantification method is integrated into Celero DNA-Seq and Universal Plus mRNA-seq library preparation kits
  • Get more consisitent results as NuQuant library quantification requires no serial dilution of standards

How do I use NuQuant?

NGS NuQuant library quantification can be performed with a simple fluorescent measurement and is compatible with Qubit and standard fluorescent plate readers. NuQuant library quantification app for Qubit can be downloaded on Github. Molar concentrations of libraries are determined by a fluorescence measurement at 650/670 nm, allowing high-throughput quantification with standard plate readers.

NuQuant library quantification workflow. After final purification of libraries using Celero™ DNA-Seq or Universal Plus mRNA-Seq with NuQuant kits, the libraries and standards are diluted in the NuQuant Library Quantifucation Buffer. The standards and libraries are assayed on the Qubit or other compatible fluorescence plate readers. The molar concentration of each library is provided for use in pooling. The pooled libraries are ready for sequencing on Illumina NGS platforms.

 

The following DNA-Seq and RNA-Seq library prepeation kits come standard with NuQuant library quantification method

ProductPart Number
Celero DNA-Seq0360-08, 0360-24
Celero DNA-Seq Metaplex0360A-96
Celero DNA-Seq + UDI0360A-UDI
Universal Plus mRNA-Seq with NuQuant0520-24, 0520-A01
Universal Plus mRNA-Seq with NuQuant, Human Globin AnyDeplete0521-24, 0521-A01