Normalization is the process by which NGS libraries of different concentrations are diluted to the same final concentration before pooling and sequencing. This step is needed to obtain best data quality from a run. Traditional normalization requires library quantification, calculations, and adjusting the concentration of each library by pipetting different volumes of buffer. Though normalization is crucial, the tradition protocol places time and cost constraints on labs – particularly those feeling the strain of workforce shortages. PerkinElmer’s new, patent-pending, bead-based normalization protocol eliminates these constraints.
With PerkinElmer’s bead-based normalization protocol, you no longer need to quantify and dilute NGS libraries prior to pooling. During this patent-pending normalization process, DNA-binding beads are added to all the libraries. A fixed amount of DNA binds to these beads. The DNA is then eluted, and the resulting libraries have approximately the same concentration even when differing inputs are used.
Choosing bead-based normalization for NGS has many benefits including reducing turnaround times and saving costs. Bead-based normalization is faster and less labor-intensive than traditional normalization methods. In addition, traditional normalization requires reagents, tubes, tips, and microwell plates, that are not needed when using beads reducing the cost of normalization. By incorporating library prep kits with PerkinElmer’s normalization beads into their workflows, laboratories can save up to three hours per 96 samples, which increases processing capacity and throughput. If the lab can process a higher number of samples, this will yield a bigger time-saving benefit for them and, in turn, decrease the cost per sample.
This process also allows normalization to be completely automated on the liquid handler the library is constructed on, further simplifying library prep, reducing human error, and increasing laboratory capacity.
The main advantage of normalization beads are the savings in reagent and consumable costs, but by also reducing the time required to load samples into the sequencer and the ability to completely automate the normalization process, normalization beads allow laboratories to process more samples.