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  • Key
  • Problem
  • Identifier
  • Solution
Identifier
  • Protein contamination
Solution
  • Re-purify samples using proteinase
Low OD 260/280 values (<1.8) Image
Identifier
  • RNA contamination
Solution
  • Ensure that your DNA extraction protocol includes RNase
High OD 260/280 values (>2.0) Image
Identifier
  • Salt or solvents contamination (e.g. Phenol)
Solution
  • Re-purify samples by ethanol precipitation
Low OD 260/230 values (<1.5) Image
Identifier
  • Drift in DNA concentration during storage
Solution
  • Ensure correct DNA input concentration by Qubit® (or similar fluorometer) reading immediately before starting the protocol
Unexpected sample concentration Image
Identifier
  • FFPE damage
Solution
  • Treat samples with SureSeq™ FFPE Repair Mix*
Low DNA integrity (DNA Integrity Score <5.5) Image
Identifier
  • gDNA too concentrated
Solution
  • Measure DNA concentration and dilute samples to ~ 70 ng/μl in nuclease-free water prior to shearing or fragmentation
Viscous sample Image
NGS tips & troubleshooting

SureSeq: For Research Use Only; Not for Diagnostic Procedures. The SureSeq NGS Library Preparation Kit was jointly developed between OGT and Bioline Reagents Limited. AMPure® is a registered trademark of Beckman Coulter Inc., Dynabeads® is a registered trademark of Thermo Fisher Scientific, TapeStation® is a registered trademark of Agilent Technologies, Inc., Qubit® is a registered trademark of Molecular Probes, Inc. Covaris® is a registered trademark of Covaris, Inc. and NEBNext® and Fragmentase® are registered trademarks of New England Biolabs, Inc. *The SureSeq FFPE DNA Repair Mix can only be purchased in conjunction with SureSeq NGS panels, not as a standalone product.

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