We are working on HBV quantification by using Rotor gene with Artus HBV PCR kit, the problem here is we are trying to work in 1/4 of the recommended volume and it is not showing similar results with the full volume results.
We have taken 7.5 microliters of master mix and 5 microliter of Extracted DNA and the recommended volume is 30 microliter of master mix and 20 microliter of extracted DNA
The results shows as follows.
For 12.5 microliter volume reaction copies are 89180253 Iu/ml.
For 50 microliter reaction copies are 166873792 Iu/ml.
What our idea is if you add 4 volumes greated amout of DNA in the 50 microliter reaction we are expecting 12.5 reaction copies * 4..But as per our results it is showing only less thandouble amount of copies
Please let us know the problem and solution.