Dear colleagues, I need support with my work in which I use the qPCR method.
In general, in my experiments, I try to measure the level of induction of prophages in different culture conditions ( in the presence of mitomycin C, different pH, temperature etc.). For this purpose, I designed some primer sets for attP (which help me to observe the number of free phages) and attB (for genomes without prophages). Additionally, to perform quantitation I designed another set of primers for endogenous control which is selected gene in a bacterial genome. The main problem is the reaction efficiency because for all primers sets I get about 80 – 90% PCR efficiency using serial dilutions of mix DNA (whole bacterial DNA + phage DNA) from all samples tested. The question is whether the results I obtained are reliable and suitable for publication. In my opinion, I observe the biological process of phage induction, however, I don't know if my results are scientifically valuable.
Thank you in advance,
Piotr Jarocki
Hello, Piotr Jarocki
You can try using "The MIQE Guidelines", which contains Minimum Information for Publication of Quantitative Real-Time PCR Experiments.
I don't know if it is your case, but it may be helpful.
Best regards,
Victoria
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