Hey,
I try ti explain as best as possible, while being relatively new to this field.
I want to perform a qPCR with the mcrA gene sequence (0.5kbp roughly) as standard. Ergo I want to quantify mcrA gene abundances in environmental samples.
As in my lab noone works on this, I order a strain with this gene (1 gene copy per organism), extracted its DNA and amplified the mcrA gene with PCR. Same day, I stained and counted the cells/ genes per µl.
My question is: when using the original strains DNA amplicons to make a qPCR Standard myself, how do I come from the strains yield to gene copies/ µl of the amplicon's yield? I am aware of the dillutions I need to perform for the eventual qPCR, but I am not clear yet on how to calculate the gene copies per µl amplicon template.
I hope I explained my point understandably, if not, I am happy to clarify.
Thanks a lot in advance
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