Tips include what to do following denaturation at 99°C, how to avoid evaporation from PCR tubes, and advice when adding reagents to a sample.
Always use a closed thermalcycler with heated Lid (105°C).
Mike Ornelas
FAS Manager – North America
Following denaturation at 99°C for 20 minutes, immediately snap chill samples on wet ice or cold blocks. Do not leave in thermalcycler to ramp down to 4°C.
Pete Gray
Array and NGS Field Application Specialist, Europe
Take care when pipetting the viscous Klenow enzyme.
Mike Ornelas
FAS Manager – North America
Avoid evaporation from PCR tubes by ensuring a good seal on caps.
Pete Gray
Array and NGS Field Application Specialist, Europe
When adding reagents to the sample, ensure efficient mixing either by rapid pipetting or gentle vortexing. Failure to mix the reagents and samples properly could lead to inefficient sample labelling and dye incorporation or uneven hybridisation of the labelled target to the arrays.
Mike Ornelas
FAS Manager – North America
Ensure all reagents are defrosted and mixed before use. To ensure all reagents are at correct concentrations prior to use they should be thoroughly thawed but not above room temperature, efficiently mixed and then gently spun to collect contents at the bottom of tube.
Pete Gray
Array and NGS Field Application Specialist, Europe
Check clean up columns are spun at the correct centrifuge speed - 2000g for 1 minute.
Mike Ornelas
FAS Manager – North America
Add labelled DNA to the centre of the gel matrix in your purification plates or columns to ensure efficient purification.
Pete Gray
Array and NGS Field Application Specialist, Europe
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