What QC steps do you recommend following extraction of DNA and prior to library preparation?

In all cases it is important to accurately determine the DNA concentration in your sample, especially when using <100 ng of DNA as input. To do this we recommend using a fluorometric method (such as the Qubit assay, Invitrogen).  Additionally, you may want to perform different QC steps on your input material according to your material type.

a) FFPE derived DNA may be compromised due to formalin fixation so we recommend performing a quality check by running your samples on a genomic screen tape (TapeStation) to determine the DIN (DNA integrity) score. This allows you to optimise the DNA input adding more material with lower DIN scores.

b) DNA derived from plasma may be contaminated with gDNA and the extent of this can be determined using a High Sensitivity D1000 Screen Tape on the TapeStation. Contamination with gDNA can arise during the processing of plasma due to haemolysis of white blood cells. A peak at >1500 bp is evidence of sample contamination with gDNA. You may also want to run this check to determine that the TapeStation profile reveals fragments of the expected size range (see Fig. 2 below).

Fig.2. Example of cell free DNA profile for patient with an inflammatory condition.