Baseline and threshold adjustment - (May/27/2005 )

Should I adjust baseline for each different target in the same run or is one baseline adjustment is enough for all different targets in the same run? How about threshold adjustment? Should I adjust threshold for each different target in the same run or is one threshold adjustment is enough for all different targets in the same run?

I don't know which system you use. If it is possible, for the reactions using the same primerpair set a baseline for them.

For setting thresfold, if you have a dilution serie as stadard curve, put it to the log phase of the curves, to the level that the distance between the curve of each dilution is the same.
Also, all curves of the samples which have the same Tm (the Tm of your amplicon) must cross the threshold, and all curves with different Tm you should avoid.

Succes!