I work with the detection of N. caninum DNA in milk samples in milk samples. The DNA concentrations in my samples range between 2-90 ng / ul. I would like to establish and standardize a final DNA concentration for my PCR reaction without having to add a very large amount of template. How much DNA template I should use?
You can buy a reference strain DNA and serially dilute to find a good working range but in real life all of the dna that you are purifying may not be N caninum so it may be difficult to standardise the amount of dna. You could mix dna samples that you have most of for this preliminary experiment to save using all of your dnaTry a series of sample from 20ng to 2 ng at 35cycles to get a rough idea of what gives you a good chance of success. the attached may be useful
Hello Dear Paul Rutland.
is this method useful for any samples such as biological samples (e.g. human serum)?
best regards
If you mean sample mixing Fatemeh Yaghoobiadeh it is often useful to have a dna stock that is a constant factor in pcr and troubleshooting and it gets round 2 problems of not enough dna and then having to change to another dna and possibly running tests on a dna sample that is poor quality and fails but now you do not know if the dna or the test conditions are the cause of the failure. Clinical geneticists may have ethical problems with sample mixing but it can be useful
- Badges
- Science method