NGS Success - Hybridisation Temperature Optimisation

View next generation sequencing (NGS) tips for the hybridisation temperature optimisation:

  • Washes must be pre-warmed as per the handbook. 
  • Test thermocyclers and consumables to be used during hybridisation and wash steps if using alternatives to minimise evaporation. Evaporation of the wash buffers alters the ratio of buffer:sample. This may lead to less efficient washing of the DNA hybridised to the streptavidin beads.

Fig. 1. Comparison of Hybridisation Wash temperatures Figure 1. Comparison of Hybridisation Wash temperatures showing minimum coverage over the 7 gene targets in the Ovarian panel. Washing at  40˚C gives the best results.

 

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