RT-PCR Result: Lower Amplification

Low amplification plots may be caused by;

• Low quality PCR strip tubes, optical tubes, plates or sealing may be used
• Master mix may be melted many times (more than 3 times), efficiency may be down
• Kits may be out of date, please check the expiration dates of the RNA isolation solutions & diagnostic kits
• There is a low amplification plots above, if there is no amplification (if there is only a single straight line on the screen) then you can perform the instrument ‘self test’ function.

Optimal Software ‘X’ (Cycle axis) and ‘Y’ (Fluorescent axis) scale adjustment may be a solution to see the amplification plots higher;

• Select ‘y‘ axis (Fluorescent value axis, when you select it correctly then the color changes to ‘blue‘
• Then use your mouse wheel to increase/decrease the scale of the ‘y’ axis.
• Amplification curves become more clear to interpret the results.

This will help you to check whether there is a sigmoidal application curve, or not. As above example, it’s clear that all internal controls are amplified as expected and there is no need to repeat any sample for this run.

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