What Sample Quality Do I Need for Library Prep?

It's important to have a high-quality DNA or RNA sample before starting the library preparation protocol.

Always follow the recommendations of your DNA or RNA isolation kit manufacturer or protocol to get the best quality possible.

Some types of input material are inherently challenging for the generation of high quality DNA or RNA, for example, DNA or RNA derived from Formalin-Fixed Paraffin-Embedded (FFPE) tissue specimens. You may need to do additional input controls:

• Evaluate the size distribution DV₂₀₀ (the percentage of fragments >200 nucleotides) of your FFPE RNA samples. This can be done using a Bioanalyzer or Fragment Analyzer from Agilent.

• Calculate the Genomic Quality Number (GQN) for every sample using the Fragment Analyzer from Agilent.

• Perform qPCR-based methods for quantitation of the library, or for presence of genes of interest or control genes.

• Alternatively, for RNA samples, Customers can obtain the RNA Integrity Number (RIN) using a Bioanalyzer

Low quality samples may need higher input amounts in library prep or sequencing, or more cycles of amplification during library prep. Final libraries may also be shorter.

For more information, see the G4 Best Practices and Quality Control Guide, reach out to your Field Application Scientist, or contact Customer Care.

October 2023