1. Nonacus Support
  2. Cell3™ Target
  3. Cell3™ Target Library preparation: quality controls (QC) and troubleshooting

My TapeStation trace shows a peak at ~160bp representing adapter-dimers, what should I do?

A small amount of adapter-dimers seen at the pre-capture library preparation QC (as seen in fig.3 Q16 above) should not be carried over to the final library pool. If your libraries show a significant adapter-dimer peak (see fig. 4 below) this may be because you have input less than 50ng of DNA into the initial reaction without diluting the adapters to 1.5 µM (1:10 dilution). Always ensure you have accurately determined the concentration in your samples immediately prior to library preparation (see Q.1). You can attempt to reduce the adapter-dimer peak by performing a 0.9X (bead to sample ratio) clean-up step. However, this will result in sample loss and you will need to perform additional PCR cycles to ensure you have sufficient material to continue.

 

adapter-dimers

Figure 4. Library preparation from cfDNA showing significant adapter-dimer peak at 152bp