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.
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.