NGS Success - Pre-Analytical QC

View next generation sequencing (NGS) tips for pre-analytical QC:

DNA Integrity: Tape Station

  • Flick Screen Tape to eliminate bubbles.
  • If insufficient sample migration occurs, dilute samples to a lower buffer salt concentration.

Fig 1. Illustrating the differences in quality of input DNA

Figure 1 - Illustrating the differences in quality of input DNA. Blue trace is poor quality FFPE sample with a DIN of 2.3. Green trace is good quality  FFPE sample with a DIN score of 5.6. Orange trace is a good quality whole blood DNA sample with a DIN score of 6.5.

Concentration: Qubit

  • Bring all reagents to room temperature.
  • Protect reagents and working solution from light.
  • Do not hold assay tubes in hand before measurement.
  • Follow recommended incubation times.
  • If your reading is too high, dilute samples.
  • If you reading is too low, protect from light which diminishes sample signal and check standard tube readings for accuracy.

Purity: Nanodrop

  • Carryover contamination may decrease or increase readings, look for contamination patterns in graph.

 

SureSeq™: For research use only; not for use in diagnostic procedures.