Dear friends and colleagues,
I have some questions, it might be stupid so forgive me in advance. I have a protocol that is used to quantify a virus, that is being extracted from plasma or serum. Before adding the extracted sample to the PCR master mix, it is asking to dilute the sample to 5ng/ul or if the concentration is not known just dilute it to 1:20. Now my questions are:
1- since we are extracting sample from serum and plasma which is considered as cell-free. How much usually the is the DNA quantity in those type of samples?
2- is it really critical to dilute the sample. I know too much DNA might inhibit PCR reaction, but given we are extracting from plasma or serum, is this critical?
3- If we did dilute the sample, dont we have to multiply the result by the dilution factor which is 20?. No such thing is mentioned in the protocol? I just need to double check.
Thanks in advance