yes chris.bailey and naavis are correct, for proper calibration the images have to be opened as raw monochrome files. debayering does pixel interpolation which invents data for the missing pixels of each color, making correct calibration impossible. your calibration frames need to be opened as raw files as well.
a manual flow would be:
BatchFormatConversion of bias sub frames (in raw mode), then integrate the .fits of the bias frames with no weighting ---> masterBias.
ImageCalibration of dark sub frames with masterBias only (this converts them to .fits as a side effect)
ImageIntegration of calibrated dark subs, no weighting, use pixel rejection to get rid of cosmic ray hits. carefully study rejection maps. --> masterDark_c
ImageCalibration of flat sub frames with masterBias only (if flat exposures are short enough)
ImageIntegration of calibrated flat frames (see vicent's flat frame tutorial for hints about how to integrate) --> masterFlat_c
ImageCalibration of light subs with masterBias, masterDark_c, masterFlat_c. since the master dark and flat are already calibrated, do not check "calibrate" under the master dark and master flat sections. the masterDark_c can be optimized (scaled) if necessary.
ImageIntegration of light subs.
the flow is (raw) open, calibrate, debayer (->rgb) star alignment, integration.
given all that stuff above, you can see why the BatchPreProcessing script was invented. for mono cameras, the flats have to be done 4 or more times depending on how many filters you have used. on the other hand, depending on how quickly dust motes move around on your sensor (and how fast your optics are), you may be able to get away with the same flats for a while. with my f/2.8 lens i can not see any dust motes in the flats and so they are good for a long time. at f/8 with my telescope, it's a different story. i need new flats for every night's work.
rob