Hi,
I would like to detect E.Coli after amplification of 16S rRNA (I am talking about part of the sequence, i.e. 230 nt). So I let cells to grow and then use Qiagen DNA/RNA extraction kit. Then I quantify what I get after extraction, it is 100 ng/µl... of what? I assume all nucleic acids, including 16S rRNA, add their "weight" to the obtained concentration of the sample.
Now suppose that I repeat experiment (I mean grow another cell line of the same strain and extract the whole nucleic acid content from there) and get the new sample concenteration 100 ng/µl. Can I really compare them? Isn't it the case that one cell line contained different amount of ribosomes than another cell line?
I have read that number of ribosomes can vary significantly depending on something like "dividing time" or "doubling time" of cells, so does anyone have a nice idea how to treat my samples? Thank you.
In order to amplify the 16SrRNA regions, you have to perform PCR reaction for whole genomic DNA and amplify it using special primers like 518F, 800R . I generally use these primers for my research.
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