I want to quantify the quantity of bifidobacterium in a milk sample. I have extracted DNA from bifidobacteria to be used as my standard, i did a 10-fold dilution and ran my standards with my unknown sample. However i need help with interpreting my results. How will i represent my results? I'm i suppose to have serially diluted my standard and culture the dilutions before running my qPCR?
You'll need to make a standard curve by diluting your standard, to get Ct values for different concentrations. Then you should be able to compare your unknown to your standard curve.
Essentially:
Run a total DNA extraction on your sample, and on your Bifidobacterium standard,
Serially dilute the DNA from your standard to create a standard curve
Compare the Ct from your unknown sample to the standard curve to find out the amount of Bifidobacterium in your unknown sample.
Compare the concentration of Bifidobacterium DNA in your unknown to a total DNA quantification by another method, and you should then see the relative abundance of Bifidobacterium in your sample.
I recommend the Robert answer, the only point that I want to mention her is that the quantification will be for both viable and dead bacteria.
Standard count culture method by using proper selective culture media for bifidobacterium in anaerobic condition is the best way for enumeration that could able to count viable bacteria.@
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