I have two sets of qPCR data taken from the same sample type at two different points in time: one was acquired by researcher A two years ago with parameters A (Ta = 57 ºC, etc.); the other by researcher B recently with parameters B (T = 59 ºC, etc.). The data compare expression levels of a single gene between wild-type and transgenic animals. Reconciliation of these two sets would avoid further animal experimentation and increase sample size. Is there any way I can peacefully do this without feeling guilty? Thanks.
Bruno Moraes As far as you sure about the specificity of primers (i.e. you are getting only one peak in melting curve reaction), you can combine the data after normalising them with internal control genes analysed at respective time.
It is likely that you would get only one band at 57 C as well as at 59 C, if primers are very much specific.
Hi Ejaj K Pathan, thank you so much for clarifying this, as it is not my area of expertise. I do now get only one peak in melting curve analysis but I have to check if my colleagues have gotten so in the past. Hopefully they did and your answer saves a few lives. Again, much appreciated.
- Badges
- Science method