Error #5: Adding reagents incorrectly
If a sample does not receive all the correct reagents in the correct order the library preparation will fail (Table 1). Some mistakes which can be made include skipping a sample when adding reagents, adding the reagents in the incorrect order, and adding the wrong reagent volumes.
Table 1. This example shows the final library yield when sample four was accidently skipped when adding the adapter ligation mastermix. The adapters were unable to ligate, preventing this sample from being amplified and generating libraries during amplification.