Tips include how to dispense hybridisation mix to the centre of the gasket chamber, why you should monitor the temperature of the hybridisation oven, and why it is essential to dislodge stuck bubbles in the hybridisation chamber.
Take extra care when pipetting the viscous hybridisation buffer.
Mike Ornelas
FAS Manager – North America
Insufficient volumes of the hybridisation mix will result in black holes on the subarrays.
Pete Gray
Array and NGS Field Application Specialist, Europe
Slowly dispense hybridisation mix to the centre of the gasket chamber. Carefully move the pipette tip around the centre to spread hybridisation mix evenly. Avoid going close to edges of gasket.
Mike Ornelas
FAS Manager – North America
Monitor the temperature of the hybridisation oven using a calibrated reference thermometer. Oven temperatures can drift over time and lead to sub-optimal hybridisation conditions.
Pete Gray
Array and NGS Field Application Specialist, Europe
Avoid scratching or cracking microarray slides by ensuring that the microarray “jig”, which holds the slides together during hybridisation, is clean and rust-free before tightening.
Mike Ornelas
FAS Manager – North America
Ensure that bubbles in the hybridisation chamber are all moving freely. Tap on hand or bench to dislodge stuck bubbles. Free movement of the bubbles is required to ensure efficient hybridisation. Any bubbles that are stuck will result in no hybridisation under the bubble and a dark hole will be seen on the scanned images.
Pete Gray
Array and NGS Field Application Specialist, Europe
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