Hello, am working with tomato RNA virus but i have been challenged by RTPCR amplification with no product using promega's access RTPCR kit. however i have been able to amplify the tomato gene in some of the samples.
Could anybody be having any remedies to this challenge of no PCR product yet results on the total RNA extracted are good and show that the samples are not degraded
Just for clarification - What is your evidence for the presence of the virus? Also, do you have any feel for the copy number?
Is it possible that the amount of virus nucleic acid is either very low, or very high, such that the amount you use in your reactions is not optimal? You can try starting with a relatively large amount of material and then doing a dilution series. Sometimes the product will appear only at low starting material concentrations.
Do you have any of the purified viral RNA you can use as a positive control?