> calibrate with bias subtracted dark frame only
Maybe I misunderstood you, but if you only subtract a bias-subtracted dark frame (or a bias-subtracted MasterDark) from the lights, the calibration result will be not correct (see:
https://pixinsight.com/forum/index.php?topic=11968 ).
A MasterDark shall be made by integration of NOT bias-subtracted darks. The MasterDark shall NOT be bias-subtracted as well. Then there are two posibilities in the calibration of the light frames:
1) Without Dark frame optimization:
Be shure to uncheck the 'Master Bias' section. In the 'Master Dark' section select the MasterDark, uncheck both 'Calibrate' and 'Optimize',
or
2) With Dark frame optimization:
Be shure to check the 'Master Bias' section and select the MasterBias. In the 'Master Dark' section select the MasterDark, check both 'Calibrate' and 'Optimize'.
If then the problem persists, try a different debayer algorithm (e.g. AHD).
Bernd